Muscarinic agonists

ABSTRACT

This invention relates to compounds that are agonists of the muscarinic M1 receptor and/or M4 receptor and which are useful in the treatment of muscarinic M1/M4 receptor mediated diseases. Also provided are pharmaceutical compositions containing the compounds and the therapeutic uses of the compounds. Compounds include those according to formula (1a)or a salt thereof, wherein p, q, r, s, Q, R3 and R4 are as defined herein.

This application is a continuation of U.S. patent application Ser. No.15/699,674, filed Sep. 8, 2017, which is a divisional of U.S. patentapplication Ser. No. 15/227,325, filed Aug. 3, 2016, now U.S. Pat. No.9,758,506, which claims priority to GB Application No.: 1513742.5, filedAug. 3, 2015. The entire contents of these applications are incorporatedherein by reference.

This invention relates to compounds that are agonists of the muscarinicM₁ receptor and/or M₄ receptor and which are useful in the treatment ofmuscarinic M₁/M₄ receptor mediated diseases. Also provided arepharmaceutical compositions containing the compounds and the therapeuticuses of the compounds.

BACKGROUND OF THE INVENTION

Muscarinic acetylcholine receptors (mAChRs) are members of the Gprotein-coupled receptor superfamily which mediate the actions of theneurotransmitter acetylcholine in both the central and peripheralnervous system. Five mAChR subtypes have been cloned, M₁ to M₅. The M₁mAChR is predominantly expressed post-synaptically in the cortex,hippocampus, striatum and thalamus; M₂ mAChRs are located predominantlyin the brainstem and thalamus, though also in the cortex, hippocampusand striatum where they reside on cholinergic synaptic terminals(Langmead et al., 2008 Br J Pharmacol). However, M₂ mAChRs are alsoexpressed peripherally on cardiac tissue (where they mediate the vagalinnervation of the heart) and in smooth muscle and exocrine glands. M₃mAChRs are expressed at relatively low level in the CNS but are widelyexpressed in smooth muscle and glandular tissues such as sweat andsalivary glands (Langmead et al., 2008 Br J Pharmacol).

Muscarinic receptors in the central nervous system, especially the M₁mAChR, play a critical role in mediating higher cognitive processing.Diseases associated with cognitive impairments, such as Alzheimer'sdisease, are accompanied by loss of cholinergic neurons in the basalforebrain (Whitehouse et al., 1982 Science). In schizophrenia, whichalso has cognitive impairment as an important component of the clinicalpicture, mAChR density is reduced in the pre-frontal cortex, hippocampusand caudate putamen of schizophrenic subjects (Dean et al., 2002 MolPsychiatry). Furthermore, in animal models, blockade or damage tocentral cholinergic pathways results in profound cognitive deficits andnon-selective mAChR antagonists have been shown to inducepsychotomimetic effects in psychiatric patients. Cholinergic replacementtherapy has largely been based on the use of acetylcholinesteraseinhibitors to prevent the breakdown of endogenous acetylcholine. Thesecompounds have shown efficacy versus symptomatic cognitive decline inthe clinic, but give rise to dose-limiting adverse events resulting fromstimulation of peripheral M₂ and M₃ mAChRs including disturbedgastrointestinal motility, bradycardia, nausea and vomiting(www.drugs.com/pro/donepezil.html; www.drugs.com/pro/rivastigmine.html).

Further discovery efforts have targeted the identification of direct M₁mAChR agonists with the aim of inducing selective improvements incognitive function with a favourable adverse effect profile. Suchefforts resulted in the identification of a range of agonists,exemplified by compounds such as xanomeline, AF267B, sabcomeline,milameline and cevimeline. Many of these compounds have been shown to behighly effective in pre-clinical models of cognition in both rodentsand/or non-human primates. Milameline has shown efficacy versusscopolamine-induced deficits in working and spatial memory in rodents;sabcomeline displayed efficacy in a visual object discrimination task inmarmosets and xanomeline reversed mAChR antagonist-induced deficits incognitive performance in a passive avoidance paradigm.

Alzheimer's disease (AD) is the most common neurodegenerative disorder(26.6 million people worldwide in 2006) that affects the elderly,resulting in profound memory loss and cognitive dysfunction. Theaetiology of the disease is complex, but is characterised by twohallmark brain pathologies: aggregates of amyloid plaques, largelycomposed of amyloid-β peptide (Aβ), and neurofibrillary tangles, formedby hyperphosphorylated tau proteins. The accumulation of Aβ is thoughtto be the central feature in the progression of AD and, as such, manyputative therapies for the treatment of AD are currently targetinginhibition of Aβ production. Aβ is derived from proteolytic cleavage ofthe membrane bound amyloid precursor protein (APP). APP is processed bytwo routes, nonamyloidgenic and amyloidgenic. Cleavage of APP byγ-secretase is common to both pathways, but in the former APP is cleavedby an α-secretase to yield soluble APPα. However, in the amyloidgenicroute, APP is cleaved by β-secretase to yield soluble APPβ and also Aβ.In vitro studies have shown that mAChR agonists can promote theprocessing of APP toward the soluble, non-amyloidogenic pathway. In vivostudies showed that the mAChR agonist, AF267B, altered disease-likepathology in the 3×TgAD transgenic mouse, a model of the differentcomponents of Alzheimer's disease (Caccamo et al., 2006 Neuron). ThemAChR agonist cevimeline has been shown to give a small, butsignificant, reduction in cerebrospinal fluid levels of Aβ inAlzheimer's patients, thus demonstrating potential disease modifyingefficacy (Nitsch et al., 2000 Neurol).

Preclinical studies have suggested that mAChR agonists display anatypical antipsychotic-like profile in a range of pre-clinicalparadigms. The mAChR agonist, xanomeline, reverses a number of dopaminemediated behaviours, including amphetamine induced locomotion in rats,apomorphine induced climbing in mice, dopamine agonist driven turning inunilateral 6-OH-DA lesioned rats and amphetamine induced motor unrest inmonkeys (without EPS liability). It also has been shown to inhibit A10,but not A9, dopamine cell firing and conditioned avoidance and inducesc-fos expression in prefrontal cortex and nucleus accumbens, but not instriatum in rats. These data are all suggestive of an atypicalantipsychotic-like profile (Mirza et al., 1999 CNS Drug Rev). Muscarinicreceptors have also been implicated in the neurobiology of addicition.The reinforcing effects of cocaine and other addictive substances aremediated by the mesolimbic dopamine system where behavioral andneurochemical studies have shown that the cholinergic muscarinicreceptor subtypes play important roles in regulation of dopaminergicneurotransmission. For example M(4) (−/−) mice demonstratedsignificantly enhanced reward driven behaviour as result of exposure tococaine (Schmidt et al Psychopharmacology (2011) August; 216(3):367-78).Furthermore xanomeline has been dmoenstrated to block the effects ofcocaine in these models.

Muscarinic receptors are also involved in the control of movement andpotentially represent novel treatments for movement disorders such asParkinson's disease, ADHD, Huntingdon's disease, tourette's syndrome andother syndromes associated with dopaminergic dysfunction as anunderlying pathogenetic factor driving disease.

Xanomeline, sabcomeline, milameline and cevimeline have all progressedinto various stages of clinical development for the treatment ofAlzheimer's disease and/or schizophrenia. Phase II clinical studies withxanomeline demonstrated its efficacy versus various cognitive symptomdomains, including behavioural disturbances and hallucinationsassociated with Alzheimer's disease (Bodick et al., 1997 Arch Neurol).This compound was also assessed in a small Phase II study ofschizophrenics and gave a significant reduction in positive and negativesymptoms when compared to placebo control (Shekhar et al., 2008 Am JPsych). However, in all clinical studies xanomeline and other relatedmAChR agonists have displayed an unacceptable safety margin with respectto cholinergic adverse events, including nausea, gastrointestinal pain,diahorrhea, diaphoresis (excessive sweating), hypersalivation (excessivesalivation), syncope and bradycardia.

Muscarinic receptors are involved in central and peripheral pain. Paincan be divided into three different types: acute, inflammatory, andneuropathic. Acute pain serves an important protective function inkeeping the organism safe from stimuli that may produce tissue damage;however management of post-surgical pain is required. Inflammatory painmay occur for many reasons including tissue damage, autoimmune response,and pathogen invasion and is triggered by the action of inflammatorymediators such as neuropeptides and prostaglandins which result inneuronal inflammation and pain. Neuropathic pain is associated withabnormal painful sensations to non-painful stimuli. Neuropathic pain isassociated with a number of different diseases/traumas such as spinalcord injury, multiple sclerosis, diabetes (diabetic neuropathy), viralinfection (such as HIV or Herpes). It is also common in cancer both as aresult of the disease or a side effect of chemotherapy. Activation ofmuscarinic receptors has been shown to be analgesic across a number ofpain states through the activation of receptors in the spinal cord andhigher pain centres in the brain. Increasing endogenous levels ofacetylcholine through acetylcholinesterase inhibitors, direct activationof muscarinic receptors with agonists or allosteric modulators has beenshown to have analgesic activity. In contrast blockade of muscarinicreceptors with antagonists or using knockout mice increases painsensitivity. Evidence for the role of the M₁ receptor in pain isreviewed by D. F. Fiorino and M. Garcia-Guzman, 2012.

More recently, a small number of compounds have been identified whichdisplay improved selectivity for the M₁ mAChR subtype over theperipherally expressed mAChR subtypes (Bridges et al., 2008 Bioorg MedChem Lett; Johnson et al., 2010 Bioorg Med Chem Lett; Budzik et al.,2010 ACS Med Chem Lett). Despite increased levels of selectivity versusthe M₃ mAChR subtype, some of these compounds retain significant agonistactivity at both this subtype and the M₂ mAChR subtype. Herein wedescribe a series of compounds which unexpectedly display high levels ofselectivity for the M₁ and/or M₄ mAChR over the M₂ and M₃ receptorsubtypes.

THE INVENTION

The present invention provides compounds having activity as muscarinicM₁ and/or M₄ receptor agonists. More particularly, the inventionprovides compounds that exhibit selectivity for the M₁ receptor and/orthe M₄ receptor relative to the M₂ and M₃ receptor subtypes.

Accordingly, in one embodiment (Embodiment 1.1), the invention providesa compound of the formula (1) or formula (1a):

or a salt thereof, wherein

p is 1 or 2;

q is 0, 1 or 2;

r is 1 or 2;

s is 0 or 1, where the total of r and s is 1 or 2;

Q is CR¹R²NR⁵R⁶, NR⁵R⁶, OR⁷, SR⁷;

R¹ is selected from hydrogen or a C₁₋₆ non-aromatic hydrocarbon groupwhich is optionally substituted with one to six fluorine atoms andwherein one or two, but not all, carbon atoms of the hydrocarbon groupmay optionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof;

R² is selected from hydrogen or a C₁₋₆ non-aromatic hydrocarbon groupwhich is optionally substituted with one to six fluorine atoms andwherein one or two, but not all, carbon atoms of the hydrocarbon groupmay optionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof;

R³ is selected from hydrogen; fluorine; cyano; hydroxy; amino; and aC₁₋₆ non-aromatic hydrocarbon group which is optionally substituted withone to six fluorine atoms and wherein one, two or three, but not all,carbon atoms of the hydrocarbon group may optionally be replaced by aheteroatom selected from O, N and S and oxidized forms thereof;

R⁴ is a hydrogen or a C₁₋₆ non-aromatic hydrocarbon group which isoptionally substituted with one to six fluorine atoms and wherein one ortwo, but not all, carbon atoms of the hydrocarbon group may optionallybe replaced by a heteroatom selected from O, N and S and oxidised formsthereof;

R⁵ is selected from hydroxy; OR⁷; COR⁷; COOR⁷; CH₂COR⁷; CH₂COOR⁷; a C₁₋₆non-aromatic hydrocarbon group which is optionally substituted with oneto six fluorine atoms and wherein one or two, but not all, carbon atomsof the hydrocarbon group may optionally be replaced by a heteroatomselected from O, N and S and oxidized forms thereof; and a group W orCH₂W where W is an optionally substituted 5- or 6-membered ringcontaining 0, 1, 2 or 3 heteroatoms selected from O, N and S andoxidized forms thereof;

R⁶ is selected from hydroxy; OR⁷; COR⁷; COOR⁷; CH₂COR⁷; CH₂COOR⁷; a C₁₋₆non-aromatic hydrocarbon group which is optionally substituted with oneto six fluorine atoms and wherein one or two, but not all, carbon atomsof the hydrocarbon group may optionally be replaced by a heteroatomselected from O, N and S and oxidized forms thereof; and a group W orCH₂W where W is an optionally substituted 5- or 6-membered ringcontaining 0, 1, 2 or 3 heteroatoms selected from O, N and S andoxidized forms thereof; and

R⁷ is selected from hydrogen a C₁₋₆ non-aromatic hydrocarbon group whichis optionally substituted with one to six fluorine atoms and wherein oneor two, but not all, carbon atoms of the hydrocarbon group mayoptionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof; and a group W or C₁₋₄ hydrocarbon group. W whereW is an optionally substituted 5- or 6-membered ring containing 0, 1, 2or 3 heteroatoms selected from O, N and S and oxidized forms thereof;

and the dotted line indicates an optional second carbon-carbon bond,provided that when a second carbon-carbon bond is present, then R³ isabsent.

or a salt thereof, wherein

q is 0, 1 or 2;

r is 1 or 2;

s is 0 or 1, where the total of r and s is 1 or 2;

Q is CR¹R²NR⁵R⁶, NR⁵R⁶, OR⁷, SR⁷;

R¹ is selected from hydrogen or a C₁₋₆ non-aromatic hydrocarbon groupwhich is optionally substituted with one to six fluorine atoms andwherein one or two, but not all, carbon atoms of the hydrocarbon groupmay optionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof;

R² is selected from hydrogen or a C₁₋₆ non-aromatic hydrocarbon groupwhich is optionally substituted with one to six fluorine atoms andwherein one or two, but not all, carbon atoms of the hydrocarbon groupmay optionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof;

R³ is selected from hydrogen; fluorine; cyano; hydroxy; amino; and aC₁₋₆ non-aromatic hydrocarbon group which is optionally substituted withone to six fluorine atoms and wherein one, two or three, but not all,carbon atoms of the hydrocarbon group may optionally be replaced by aheteroatom selected from O, N and S and oxidized forms thereof;

R⁴ is a hydrogen or a C₁₋₆ non-aromatic hydrocarbon group which isoptionally substituted with one to six fluorine atoms and wherein one ortwo, but not all, carbon atoms of the hydrocarbon group may optionallybe replaced by a heteroatom selected from O, N and S and oxidised formsthereof;

R⁵ is selected from hydroxy; OR⁷; COR⁷; COOR⁷; CH₂COR⁷; CH₂COOR⁷; a C₁₋₆non-aromatic hydrocarbon group which is optionally substituted with oneto six fluorine atoms and wherein one or two, but not all, carbon atomsof the hydrocarbon group may optionally be replaced by a heteroatomselected from O, N and S and oxidized forms thereof; and a group W orCH₂W where W is an optionally substituted 5- or 6-membered ringcontaining 0, 1, 2 or 3 heteroatoms selected from O, N and S andoxidized forms thereof;

R⁶ is selected from hydroxy; OR⁷; COR⁷; COOR⁷; CH₂COR⁷; CH₂COOR⁷; a C₁₋₆non-aromatic hydrocarbon group which is optionally substituted with oneto six fluorine atoms and wherein one or two, but not all, carbon atomsof the hydrocarbon group may optionally be replaced by a heteroatomselected from O, N and S and oxidized forms thereof; and a group W orCH₂W where W is an optionally substituted 5- or 6-membered ringcontaining 0, 1, 2 or 3 heteroatoms selected from O, N and S andoxidized forms thereof; and

R⁷ is selected from hydrogen a C₁₋₆ non-aromatic hydrocarbon group whichis optionally substituted with one to six fluorine atoms and wherein oneor two, but not all, carbon atoms of the hydrocarbon group mayoptionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof; and a group W or CH₂W where W is an optionallysubstituted 5- or 6-membered ring containing 0, 1, 2 or 3 heteroatomsselected from O, N and S and oxidized forms thereof;

and the dotted line indicates an optional second carbon-carbon bond,provided that when a second carbon-carbon bond is present, then R³ isabsent.

Particular compounds of the formula (1) or formula (1a) are as definedin the Embodiments 1.2 to 1.50 set out below.

1.2 A compound according to Embodiment 1.1 wherein Q is NR⁵R⁶

1.3 A compound according to Embodiment 1.1 wherein Q is CR¹R²NR⁵R⁶.

1.4 A compound according to Embodiments 1.1 to 1.3 wherein R¹ isselected from hydrogen or a C₁₋₃ alkyl group.

1.5 A compound according to Embodiment 1.4 wherein R¹ is selected fromhydrogen, methyl or ethyl.

1.6 A compound according to Embodiments 1.1 to 1.5 wherein R² isselected from hydrogen or a C₁₋₃ alkyl group.

1.7 A compound according to Embodiment 1.6 wherein R² is selected fromhydrogen, methyl or ethyl.

1.8 A compound according to Embodiment 1.6 wherein R¹ is H and R² isselected from hydrogen or methyl.

1.9 A compound according to any one of Embodiments 1.1 to 1.8 wherein R⁵is selected from a C₁₋₆ non-aromatic hydrocarbon group which isoptionally substituted with one to six fluorine atoms and wherein one ortwo, but not all, carbon atoms of the hydrocarbon group may optionallybe replaced by a heteroatom selected from O, N and S and oxidized formsthereof; and a group W or CH₂W where W is an optionally substituted 5-or 6-membered ring containing 0, 1, 2 or 3 heteroatoms selected from O,N and S and oxidized forms thereof.

1.10 A compound according to any one of Embodiments 1.1 to 1.8 whereinR⁵ is selected from a C₁₋₄ alkyl group which is optionally substitutedwith one to four fluorine atoms and wherein one or two, but not all,carbon atoms of the hydrocarbon group may optionally be replaced by aheteroatom selected from O, N and S and oxidized forms thereof; and agroup W or CH₂W where W is an optionally substituted 5- or 6-memberedaromatic ring containing 0, 1, 2 or 3 heteroatoms selected from O, N andS.

1.11 A compound according to any one of Embodiments 1.1 to 1.8 whereinR⁵ is selected from a C₁₋₄ alkyl group which is optionally substitutedwith one to four fluorine atoms; and a group W or CH₂W where W is anoptionally substituted 5- or 6-membered aromatic ring containing 0, 1, 2or 3 heteroatoms selected from O, N and S.

1.12 A compound according to any one of Embodiments 1.1 to 1.8 whereinR⁵ is selected from methyl, ethyl, propyl, isopropyl, cyclopropyl,fluoroethyl, difluoroethyl, trifluoroethyl, butyl or cyclobutyl.

1.13 A compound according to any one of Embodiments 1.1 to 1.8 whereinR⁵ is a group W or CH₂W where W is an optionally substituted phenyl,pyridyl or isoxazole ring.

1.14 A compound according to any one of Embodiments 1.1 to 1.13 whereinR⁶ is selected from; COR⁷; COOR⁷; CH₂COR⁷; CH₂COOR⁷ or a C₁₋₆non-aromatic hydrocarbon group which is optionally substituted with oneto six fluorine atoms and wherein one or two, but not all, carbon atomsof the hydrocarbon group may optionally be replaced by a heteroatomselected from O, N and S and oxidized forms thereof.

1.15 A compound according to any one of Embodiments 1.1 to 1.13 whereinR⁶ is selected from methyl, ethyl, trifluoroethyl, hydroxyethyl ormethoxyethyl.

1.16 A compound according to any one of Embodiments 1.1 to 1.13 whereinR⁶ is selected from; COR⁷; COOR⁷; CH₂COR⁷; CH₂COOR⁷, wherein R⁷ isselected from H, methyl, fluoromethyl, difluoromethyl, trifluoromethyl,ethyl, fluoroethyl, difluoroethyl or trifluoroethyl.

1.17 A compound according to any one of Embodiments 1.1 to 1.16 whereinthe dotted line represents a second carbon-carbon bond and R³ is absent.

1.18 A compound according to any one of Embodiments 1.1 to 1.16 whereinR³ is present and the optional second carbon-carbon bond is absent.

1.19 A compound according to Embodiment 1.18 wherein R³ is selected fromhydrogen; fluorine; cyano; hydroxy; amino; and a C₁₋₆ non-aromatichydrocarbon group which is optionally substituted with one to sixfluorine atoms and wherein one or two, but not all, carbon atoms of thehydrocarbon group may optionally be replaced by a heteroatom selectedfrom O, N and S and oxidized forms thereof.

1.20 A compound according to Embodiment 1.19 wherein R³ is selected fromhydrogen; fluorine; cyano; hydroxy; amino; and a C₁₋₄ alkyl group whichis optionally substituted with one to four fluorine atoms.

1.21 A compound according to Embodiment 1.20 wherein R³ is selected fromhydrogen; fluorine; cyano; hydroxy; amino; C₁₋₄ alkyl and C₁₋₄ alkoxy,wherein the C₁₋₄ alkyl and C₁₋₄ alkoxy are each optionally substitutedwith one to four fluorine atoms.

1.22 A compound according to Embodiment 1.21 wherein R³ is selected fromhydrogen; fluorine; hydroxy and methoxy.

1.23 A compound according to Embodiment 1.22 wherein R³ is hydrogen.

1.24 A compound according to any one of Embodiments 1.1 to 1.23 whereinR⁴ is hydrogen or an acyclic C₁₋₆ hydrocarbon group.

1.25 A compound according to Embodiment 1.24 wherein R⁴ is hydrogen oran acyclic C₁₋₃ hydrocarbon group.

1.26 A compound according to Embodiment 1.25 wherein R⁴ is hydrogen or aC₁₋₃ alkyl group or a C₂₋₃ alkynyl group.

1.27 A compound according to Embodiment 1.26 wherein R⁴ is selected fromhydrogen, methyl, ethyl, ethynyl and 1-propynyl.

1.28 A compound according to Embodiment 1.27 wherein R⁴ is selected fromhydrogen and methyl.

1.29 A compound according to Embodiment 1.28 wherein R⁴ is methyl.

1.30 A compound according to any one of the preceding Embodimentswherein R⁷, when present, is selected from hydrogen a C₁₋₆ non-aromatichydrocarbon group which is optionally substituted with one to sixfluorine atoms and wherein one or two, but not all, carbon atoms of thehydrocarbon group may optionally be replaced by a heteroatom selectedfrom O, N and S and oxidized forms thereof; and a group W or CH₂W orC₁₋₄ hydrocarbon group. W where W is an optionally substituted 5- or6-membered ring containing 0, 1, 2 or 3 heteroatoms selected from O, Nand S and oxidized forms thereof;

1.31 A compound according to Embodiment 1.30 wherein R⁷ is anon-aromatic C₁₋₄ hydrocarbon group optionally substituted with one ormore fluorine atoms.

1.32 A compound according to Embodiment 1.30 wherein R⁷ is a C₁₋₄ alkylgroup.

1.33 A compound according to any one of Embodiments 1.1 to 1.32 whereinq is 0.

1.34 A compound according to any one of Embodiments 1.1 to 1.32 whereinq is 1.

1.35 A compound according to any one of Embodiments 1.1 to 1.32 whereinq is 2.

1.36 A compound according to any one of Embodiments 1.1 to 1.35 whereinr is 1.

1.37 A compound according to any one of Embodiments 1.1 to 1.35 whereins is 0.

1.38 A compound according to any one of Embodiments 1.1 to 1.36 whereinr is 1 and s is 1.

1.39 A compound according to any one of Embodiments 1.1 to 1.37 whereinr is 1 and s is 0.

1.40 A compound according to any one of Embodiments 1.1 to 1.39 whereinp is 1.

1.41 A compound according to any one of Embodiments 1.1 to 1.39 whereinp is 2.

1.42 A compound according to any one of Embodiments 1.1 to 1.41 whereinthe moiety:

is selected from groups A to KKK below:

1.43 A compound according to having the formula (2) or formula (2a):

wherein p, q, r, s, R⁴R⁵ and R⁶ are is as defined in any one ofEmbodiments 1.1 to 1.39.

1.44 A compound according to having the formula (3) or formula (3a):

wherein R¹ is H or methyl and p, q, r, s, R⁴ R⁵ and R⁶ are is as definedin any one of Embodiments 1.1 to 1.41.

1.45 A compound according to having the formula (4):

wherein q is 1 or 2 and p, R⁴ R⁵ and R⁶ are is as defined in any one ofEmbodiments 1.1 to 1.32.

1.46 A compound according to Embodiment 1.1 which is as defined in anyone of Examples 1-1 to 4-1.

1.47 A compound according to any one of Embodiments 1.1 to 1.46 having amolecular weight of less than 550.

1.48 A compound according to Embodiment 1.47 having a molecular weightof less than 500.

1.49 A compound according to Embodiment 1.48 having a molecular weightof, or less than 450.

1.50 A compound according to any one of Embodiments 1.1 to 1.49 which isin the form of a salt.

1.51 A compound according to Embodiment 1.50 wherein the salt is an acidaddition salt.

1.52 A compound according to Embodiment 1.50 or Embodiment 1.51 whereinthe salt is a pharmaceutically acceptable salt.

Definitions

In this application, the following definitions apply, unless indicatedotherwise.

The term “treatment”, in relation to the uses of the compounds of theformula (1) or formula (1a), is used to describe any form ofintervention where a compound is administered to a subject sufferingfrom, or at risk of suffering from, or potentially at risk of sufferingfrom the disease or disorder in question. Thus, the term “treatment”covers both preventative (prophylactic) treatment and treatment wheremeasurable or detectable symptoms of the disease or disorder are beingdisplayed.

The term “effective therapeutic amount” as used herein (for example inrelation to methods of treatment of a disease or condition) refers to anamount of the compound which is effective to produce a desiredtherapeutic effect. For example, if the condition is pain, then theeffective therapeutic amount is an amount sufficient to provide adesired level of pain relief. The desired level of pain relief may be,for example, complete removal of the pain or a reduction in the severityof the pain.

The term “non-aromatic hydrocarbon group” as in “C₁₋₁₀ non-aromatichydrocarbon group” or “acyclic C₁₋₅ non-aromatic hydrocarbon group”refers to a group consisting of carbon and hydrogen atoms and whichcontains no aromatic rings. The hydrocarbon group may be fully saturatedor may contain one or more carbon-carbon double bonds or carbon-carbontriple bonds, or mixtures of double and triple bonds. The hydrocarbongroup may be a straight chain or branched chain group or may consist ofor contain a cyclic group. Thus the term non-aromatic hydrocarbonincludes alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,cycloalkylalkyl, cycloalkenyl alkyl and so on.

The terms “alkyl”, “alkenyl”, “alkynyl”, “cycloalkyl” aryl, heteroaryland “cycloalkenyl” are used in their conventional sense (e.g. as definedin the IUPAC Gold Book) unless indicated otherwise.

The term “saturated hydrocarbon group” as in “C₁₋₄ saturated hydrocarbongroup” refers to a hydrocarbon group containing no carbon-carbon doublebonds or triple bonds. The saturated hydrocarbon group can therefore bean alkyl group, a cycloalkyl group, a cycloalkylalkyl group, analkylcycloalkyl group or a alkylcycloalkylalkyl group. Examples of C₁₋₄saturated hydrocarbon groups include C₁₋₄ alkyl groups, cyclopropyl,cyclobutyl and cyclopropylmethyl.

The term “cycloalkyl” as used herein, where the specified number ofcarbon atoms permits, includes both monocyclic cycloalkyl groups such ascyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl, andbicyclic and tricyclic groups. Bicyclic cycloalkyl groups includebridged ring systems such as bicycloheptane, bicyclooctane andadamantane.

In the definitions of R¹, R², R³ and R⁴ above, where stated, one or twobut not all, carbon atoms of the non-aromatic hydrocarbon group mayoptionally be replaced by a heteroatom selected from O, N and S and (inthe case of R¹ and R⁴) oxidised forms thereof. It will be appreciatedthat when a carbon atom is replaced by a heteroatom, the lower valenciesof the heteroatoms compared to carbon means that fewer atoms will bebonded to the heteroatoms than would have been bonded to the carbon atomthat has been replaced. Thus, for example, replacement of of a carbonatom (valency of four) in a CH₂ group by oxygen (valency of two) willmean that the resulting molecule will contain two less hydrogen atomsand replacement of a carbon atom (valency of four) in a CH₂ group bynitrogen (valency of three) will mean that the resulting molecule willcontain one less hydrogen atom.

Examples of a heteroatom replacements for carbon atoms includereplacement of a carbon atom in a —CH₂—CH₂—CH₂— chain with oxygen orsulfur to give an ether —CH₂—O—CH₂— or thioether —CH₂—S—CH₂—,replacement of a carbon atom in a group CH₂—C≡C—H with nitrogen to givea nitrile (cyano) group CH₂—C≡N, replacement of a carbon atom in a group—CH₂—CH₂—CH₂— with C═O to give a ketone —CH₂—C(O)—CH₂—, replacement of acarbon atom in a group —CH₂—CH₂—CH₂— with S═O or SO₂ to give a sulfoxide—CH₂—S(O)—CH₂— or sulfone —CH₂—S(O)₂—CH₂—, replacement of a carbon atomin a —CH₂—CH₂—CH₂-chain with C(O)NH to give an amide —CH₂—CH₂—C(O)—NH—,replacement of a carbon atom in a —CH₂—CH₂—CH₂— chain with nitrogen togive an amine —CH₂—NH—CH₂—, and replacement of a carbon atom in a—CH₂—CH₂—CH₂— chain with C(O)O to give an ester (or carboxylic acid)—CH₂—CH₂—C(O)—O—. In each such replacement, at least one carbon atom ofthe hydrocarbon group must remain.

Salts

Many compounds of the formula (1) or formula (1a) can exist in the formof salts, for example acid addition salts or, in certain cases salts oforganic and inorganic bases such as carboxylate, sulfonate and phosphatesalts. All such salts are within the scope of this invention, andreferences to compounds of the formula (1) or formula (1a) include thesalt forms of the compounds as defined in Embodiments 1.50 to 1.52.

The salts are typically acid addition salts.

The salts of the present invention can be synthesized from the parentcompound that contains a basic or acidic moiety by conventional chemicalmethods such as methods described in Pharmaceutical Salts: Properties,Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth(Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002.Generally, such salts can be prepared by reacting the free acid or baseforms of these compounds with the appropriate base or acid in water orin an organic solvent, or in a mixture of the two; generally, nonaqueousmedia such as ether, ethyl acetate, ethanol, isopropanol, oracetonitrile are used.

Acid addition salts (as defined in Embodiment 1.120) may be formed witha wide variety of acids, both inorganic and organic. Examples of acidaddition salts falling within Embodiment 1.120 include mono- or di-saltsformed with an acid selected from the group consisting of acetic,2,2-dichloroacetic, adipic, alginic, ascorbic (e.g. L-ascorbic),L-aspartic, benzenesulfonic, benzoic, 4-acetamidobenzoic, butanoic, (+)camphoric, camphor-sulfonic, (+)-(1 S)-camphor-10-sulfonic, capric,caproic, caprylic, cinnamic, citric, cyclamic, dodecylsulfuric,ethane-1,2-disulfonic, ethanesulfonic, 2-hydroxyethanesulfonic, formic,fumaric, galactaric, gentisic, glucoheptonic, D-gluconic, glucuronic(e.g. D-glucuronic), glutamic (e.g. L-glutamic), α-oxoglutaric,glycolic, hippuric, hydrohalic acids (e.g. hydrobromic, hydrochloric,hydriodic), isethionic, lactic (e.g. (+)-L-lactic, (±)-DL-lactic),lactobionic, maleic, malic, (−)-L-malic, malonic, (±)-DL-mandelic,methanesulfonic, naphthalene-2-sulfonic, naphthalene-1,5-disulfonic,1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic,palmitic, pamoic, phosphoric, propionic, pyruvic, L-pyroglutamic,salicylic, 4-amino-salicylic, sebacic, stearic, succinic, sulfuric,tannic, (+)-L-tartaric, thiocyanic, p-toluenesulfonic, undecylenic andvaleric acids, as well as acylated amino acids and cation exchangeresins.

Where the compounds of the formula (1) or formula (1a) contain an aminefunction, these may form quaternary ammonium salts, for example byreaction with an alkylating agent according to methods well known to theskilled person. Such quaternary ammonium compounds are within the scopeof formula (1) or formula (1a) respectively.

The compounds of the invention may exist as mono- or di-salts dependingupon the pKa of the acid from which the salt is formed.

The salt forms of the compounds of the invention are typicallypharmaceutically acceptable salts, and examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1-19. However, saltsthat are not pharmaceutically acceptable may also be prepared asintermediate forms which may then be converted into pharmaceuticallyacceptable salts. Such non-pharmaceutically acceptable salts forms,which may be useful, for example, in the purification or separation ofthe compounds of the invention, also form part of the invention.

Stereoisomers

Stereoisomers are isomeric molecules that have the same molecularformula and sequence of bonded atoms but which differ only in thethree-dimensional orientations of their atoms in space. Thestereoisomers can be, for example, geometric isomers or optical isomers.

Geometric Isomers

With geometric isomers, the isomerism is due to the differentorientations of an atom or group about a double bond, as in cis andtrans (Z and E) isomerism about a carbon-carbon double bond, or cis andtrans isomers about an amide bond, or syn and anti isomerism about acarbon nitrogen double bond (e.g. in an oxime), or rotational isomerismabout a bond where there is restricted rotation, or cis and transisomerism about a ring such as a cycloalkane ring.

Accordingly, in another embodiment (Embodiment 1.121), the inventionprovides a geometric isomer of a compound according to any one ofEmbodiments 1.1 to 1.52.

Optical Isomers

Where compounds of the formula contain one or more chiral centres, andcan exist in the form of two or more optical isomers, references to thecompounds include all optical isomeric forms thereof (e.g. enantiomers,epimers and diastereoisomers), either as individual optical isomers, ormixtures (e.g. racemic mixtures) or two or more optical isomers, unlessthe context requires otherwise.

Accordingly, in another embodiment (Embodiment 1.132) the inventionprovides a compound according to any one of Embodiments 1.1 to 1.121which contains a chiral centre.

The optical isomers may be characterised and identified by their opticalactivity (i.e. as + and − isomers, or d and l isomers) or they may becharacterised in terms of their absolute stereochemistry using the “Rand S” nomenclature developed by Cahn, Ingold and Prelog, see AdvancedOrganic Chemistry by Jerry March, 4^(th) Edition, John Wiley & Sons, NewYork, 1992, pages 109-114, and see also Cahn, Ingold & Prelog, Angew.Chem. Int. Ed. Engl., 1966, 5, 385-415. Optical isomers can be separatedby a number of techniques including chiral chromatography(chromatography on a chiral support) and such techniques are well knownto the person skilled in the art. As an alternative to chiralchromatography, optical isomers can be separated by formingdiastereoisomeric salts with chiral acids such as (+)-tartaric acid,(−)-pyroglutamic acid, (−)-di-toluoyl-L-tartaric acid, (+)-mandelicacid, (−)-malic acid, and (−)-camphorsulphonic, separating thediastereoisomers by preferential crystallisation, and then dissociatingthe salts to give the individual enantiomer of the free base.

Where compounds of the invention exist as two or more optical isomericforms, one enantiomer in a pair of enantiomers may exhibit advantagesover the other enantiomer, for example, in terms of biological activity.Thus, in certain circumstances, it may be desirable to use as atherapeutic agent only one of a pair of enantiomers, or only one of aplurality of diastereoisomers.

Accordingly, in another embodiment (Embodiment 1.133), the inventionprovides compositions containing a compound according to Embodiment1.132 having one or more chiral centres, wherein at least 55% (e.g. atleast 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%) of the compound ofEmbodiment 1.108 is present as a single optical isomer (e.g. enantiomeror diastereoisomer).

In one general embodiment (Embodiment 1.134), 99% or more (e.g.substantially all) of the total amount of the compound (or compound foruse) of Embodiment 1.132 is present as a single optical isomer.

For example, in one embodiment (Embodiment 1.135) the compound ispresent as a single enantiomer.

In another embodiment (Embodiment 1.136), the compound is present as asingle diastereoisomer.

The invention also provides mixtures of optical isomers, which may beracemic or non-racemic. Thus, the invention provides:

1.137 A compound according to Embodiment 1.132 which is in the form of aracemic mixture of optical isomers.

1.138 A compound according to Embodiment 1.132 which is in the form of anon-racemic mixture of optical isomers.

Isotopes

The compounds of the invention as defined in any one of Embodiments 1.1to 1.138 may contain one or more isotopic substitutions, and a referenceto a particular element includes within its scope all isotopes of theelement. For example, a reference to hydrogen includes within its scope¹H, ²H (D), and ³H (T). Similarly, references to carbon and oxygeninclude within their scope respectively ¹²C, ¹³C and ¹⁴C and ¹⁶O and¹⁸O.

In an analogous manner, a reference to a particular functional groupalso includes within its scope isotopic variations, unless the contextindicates otherwise. For example, a reference to an alkyl group such asan ethyl group also covers variations in which one or more of thehydrogen atoms in the group is in the form of a deuterium or tritiumisotope, e.g. as in an ethyl group in which all five hydrogen atoms arein the deuterium isotopic form (a perdeuteroethyl group).

The isotopes may be radioactive or non-radioactive. In one embodiment ofthe invention (Embodiment 1.142), the compound of any one of Embodiments1.1 to 1.140 contains no radioactive isotopes. Such compounds arepreferred for therapeutic use. In another embodiment (Embodiment 1.143),however, the compound of any one of Embodiments 1.1 to 1.140 may containone or more radioisotopes. Compounds containing such radioisotopes maybe useful in a diagnostic context.

Solvates

Compounds of the formula (1) or formula (1a) as defined in any one ofEmbodiments 1.1 to 1.143 may form solvates. Preferred solvates aresolvates formed by the incorporation into the solid state structure(e.g. crystal structure) of the compounds of the invention of moleculesof a non-toxic pharmaceutically acceptable solvent (referred to below asthe solvating solvent). Examples of such solvents include water,alcohols (such as ethanol, isopropanol and butanol) anddimethylsulphoxide. Solvates can be prepared by recrystallising thecompounds of the invention with a solvent or mixture of solventscontaining the solvating solvent. Whether or not a solvate has beenformed in any given instance can be determined by subjecting crystals ofthe compound to analysis using well known and standard techniques suchas thermogravimetric analysis (TGE), differential scanning calorimetry(DSC) and X-ray crystallography. The solvates can be stoichiometric ornon-stoichiometric solvates. Particularly preferred solvates arehydrates, and examples of hydrates include hemihydrates, monohydratesand dihydrates.

Accordingly, in further embodiments 1.153 and 1.154, the inventionprovides:

1.153 A compound according to any one of Embodiments 1.1 to 1.143 in theform of a solvate.

1.154 A compound according to Embodiment 1.153 wherein the solvate is ahydrate.

For a more detailed discussion of solvates and the methods used to makeand characterise them, see Bryn et al., Solid-State Chemistry of Drugs,Second Edition, published by SSCI, Inc of West Lafayette, Ind., USA,1999, ISBN 0-967-06710-3.

Alternatively, rather than existing as a hydrate, the compound of theinvention may be anhydrous. Therefore, in another embodiment (Embodiment1.155), the invention provides a compound as defined in any one ofEmbodiments 1.1 to 1.143 in an anhydrous form (e.g. anhydrouscrystalline form).

Crystalline and Amorphous Forms

The compounds of any one of Embodiments 1.1 to 1.155 may exist in acrystalline or non-crystalline (e.g. amorphous) state. Whether or not acompound exists in a crystalline state can readily be determined bystandard techniques such as X-ray powder diffraction (XRPD). Crystalsand their crystal structures can be characterised using a number oftechniques including single crystal X-ray crystallography, X-ray powderdiffraction (XRPD), differential scanning calorimetry (DSC) and infrared spectroscopy, e.g. Fourier Transform infra-red spectroscopy (FTIR).The behaviour of the crystals under conditions of varying humidity canbe analysed by gravimetric vapour sorption studies and also by XRPD.Determination of the crystal structure of a compound can be performed byX-ray crystallography which can be carried out according to conventionalmethods such as those described herein and as described in Fundamentalsof Crystallography, C. Giacovazzo, H. L. Monaco, D. Viterbo, F.Scordari, G. Gilli, G. Zanotti and M. Catti, (International Union ofCrystallography/Oxford University Press, 1992 ISBN 0-19-855578-4 (p/b),0-19-85579-2 (h/b)). This technique involves the analysis andinterpretation of the X-ray diffraction of single crystal. In anamorphous solid, the three dimensional structure that normally exists ina crystalline form does not exist and the positions of the moleculesrelative to one another in the amorphous form are essentially random,see for example Hancock et al. J. Pharm. Sci. (1997), 86, 1).

Accordingly, in further embodiments, the invention provides:

1.160 A compound according to any one of Embodiments 1.1 to 1.155 in acrystalline form.

1.161 A compound according to any one of Embodiments 1.1 to 1.155 whichis:

(a) from 50% to 100% crystalline, and more particularly is at least 50%crystalline, or at least 60% crystalline, or at least 70% crystalline,or at least 80% crystalline, or at least 90% crystalline, or at least95% crystalline, or at least 98% crystalline, or at least 99%crystalline, or at least 99.5% crystalline, or at least 99.9%crystalline, for example 100% crystalline.

1.162 A compound according to any one of Embodiments 1.1 to 1.155 whichis in an amorphous form.

Prodrugs

The compounds of the formula (1) or formula (1a) as defined in any oneof Embodiments 1.1 to 1.162 may be presented in the form of a pro-drug.By “prodrugs” is meant for example any compound that is converted invivo into a biologically active compound of the formula (1) or formula(1a) respectively, as defined in any one of Embodiments 1.1 to 1.162.

For example, some prodrugs are esters of the active compound (e.g., aphysiologically acceptable metabolically labile ester). Duringmetabolism, the ester group (—C(═O)OR) is cleaved to yield the activedrug. Such esters may be formed by esterification, for example, of anyhydroxyl groups present in the parent compound with, where appropriate,prior protection of any other reactive groups present in the parentcompound, followed by deprotection if required.

Also, some prodrugs are activated enzymatically to yield the activecompound, or a compound which, upon further chemical reaction, yieldsthe active compound (for example, as in ADEPT, GDEPT, LIDEPT, etc.). Forexample, the prodrug may be a sugar derivative or other glycosideconjugate, or may be an amino acid ester derivative.

Accordingly, in another embodiment (Embodiment 1.170), the inventionprovides a pro-drug of a compound as defined in any one of Embodiments1.1 to 1.170 wherein the compound contains a functional group which isconvertable under physiological conditions to form a hydroxyl group oramino group.

Complexes and Clathrates

Also encompassed by formula (1) or formula (1a) in Embodiments 1.1 to1.170 are complexes (e.g. inclusion complexes or clathrates withcompounds such as cyclodextrins, or complexes with metals) of thecompounds of Embodiments 1.1 to 1.170.

Accordingly, in another embodiment (Embodiment 1.180), the inventionprovides a compound according to any one of Embodiments 1.1 to 1.170 inthe form of a complex or clathrate.

Biological Activity and Therapeutic Uses

The compounds of the present invention have activity as muscarinic M₁and/or M₄ receptor agonists. The muscarinic activity of the compoundscan be determined using the Phospho-ERK1/2 assay described in Example Abelow.

A significant advantage of compounds of the invention is that they arehighly selective for the M₁ and/or M₄ receptor relative to the M₂ and M₃receptor subtypes.

Compounds of the invention are not agonists of the M₂ and M₃ receptorsubtypes. For example, whereas compounds of the invention typically havepEC₅₀ values of at least 6 (preferably at least 6.5) and E_(max) valuesof greater than 80 (preferably greater than 95) against the M₁ receptorin the functional assay described in Example A, they may have pEC₅₀values of less than 5 and E_(max) values of less than 20% when testedagainst the M₂ and M₃ subtypes in the functional assay of Example A.

Accordingly, in Embodiments 2.1 to 2.9, the invention provides:

2.1 A compound according to any one of Embodiments 1.1 to 1.180 for usein medicine.

2.2 A compound according to any one of Embodiments 1.1 to 1.180 for useas a muscarinic M₁ and/or M₄ receptor agonist.

2.3 A compound according to any one of Embodiments 1.1 to 1.180 which isa muscarinic M₁ receptor agonist having a pEC₅₀ in the range from 6.0 to8.0 and an E_(max) of at least 90 against the M₁ receptor in the assayof Example A herein or an assay substantially similar thereto.

2.4 A compound according to Embodiment 2.3 which is a muscarinic M₁receptor agonist having a pEC₅₀ in the range from 6.5 to 7.5.

2.5 A compound according to Embodiment 2.3 or Embodiment 2.4 having anE_(max) of at least 95 against the M₁ receptor.

2.6 A compound according to any one of Embodiments 1.1 to 1.180 which isa muscarinic M₄ receptor agonist having a pEC₅₀ in the range from 6.0 to8.5 and an E_(max) of at least 90 against the M₄ receptor in the assayof Example A herein or an assay substantially similar thereto.

2.7 A compound according to Embodiment 2.6 which is a muscarinic M₄receptor agonist having a pEC₅₀ in the range from 6.5 to 8.5.

2.8 A compound according to Embodiment 2.6 or Embodiment 2.7 having anE_(max) of at least 95 against the M₄ receptor.

2.9 A compound according to any one of Embodiments 2.3 to 2.8 which isselective for the M₁ and/or M₄ receptor compared to the muscarinic M₂and M₃ receptors.

2.10 A compound according to Embodiment 2.9 which is selective for theM₁ receptor compared to the muscarinic M₂ and M₃ receptors.

2.11 A compound according to Embodiment 2.9 which is selective for theM₄ receptor compared to the muscarinic M₂ and M₃ receptors.

2.12 A compound according to any one of Embodiments 2.3 to 2.5 which isselective for the M₁ receptor compared to the muscarinic M₂, M₃ and M₄receptors.

2.13 A compound according to any one of Embodiments 2.6 to 2.8 which isselective for the M₄ receptor compared to the muscarinic M₁, M₂ and M₃receptors.

2.14 A compound according to any one of Embodiments 2.3 to 2.8 which isselective for the M₁ and M₄ receptor compared to the muscarinic M₂ andM₃ receptors.

2.15 A compound according to any one of Embodiments 2.3 to 2.14 whichhas a pEC₅₀ of less than 5 and an E_(max) of less than 50 against themuscarinic M₂ and M₃ receptor subtypes.

2.16 A compound according to Embodiment 2.15 which has a pEC₅₀ of lessthan 4.5 and/or an E_(max) of less than 30 against the muscarinic M₂ andM₃ receptor subtypes.

2.17 A compound according to any one of Embodiments 1.1 to 1.180 andEmbodiments 2.3 to 2.16 for use in the treatment of a disease orcondition mediated by the muscarinic M₁ receptor.

By virtue of their muscarinic M₁ and/or M₄ receptor agonist activity,compounds of the invention can be used in the treatment of Alzheimer'sdisease, schizophrenia and other psychotic disorders, cognitivedisorders and other diseases mediated by the muscarinic M₁ and/or M₄receptor, and can also be used in the treatment of various types ofpain.

Accordingly, in Embodiments 2.18 to 2.34, the invention provides:

2.18 A compound according to any one of Embodiments 1.1 to 1.180 for usein the treatment of a cognitive disorder or psychotic disorder.

2.19 A compound for use in according to Embodiment 2.18 wherein thecognitive disorder or psychotic disorder comprises, arises from or isassociated with a condition selected from cognitive impairment, MildCognitive Impairment, frontotemporal dementia, vascular dementia,dementia with Lewy bodies, presenile dementia, senile dementia,Friederich's ataxia, Down's syndrome, Huntington's chorea, hyperkinesia,mania, Tourette's syndrome, Alzheimer's disease, progressivesupranuclear palsy, impairment of cognitive functions includingattention, orientation, learning disorders, memory (i.e. memorydisorders, amnesia, amnesic disorders, transient global amnesia syndromeand age-associated memory impairment) and language function; cognitiveimpairment as a result of stroke, Huntington's disease, Pick disease,Aids-related dementia or other dementia states such as Multiinfarctdementia, alcoholic dementia, hypotiroidism-related dementia, anddementia associated to other degenerative disorders such as cerebellaratrophy and amyotropic lateral sclerosis; other acute or sub-acuteconditions that may cause cognitive decline such as delirium ordepression (pseudodementia states) trauma, head trauma, age relatedcognitive decline, stroke, neurodegeneration, drug-induced states,neurotoxic agents, age related cognitive impairment, autism relatedcognitive impairment, Down's syndrome, cognitive deficit related topsychosis, and post-electroconvulsive treatment related cognitivedisorders; cognitive disorders due to drug abuse or drug withdrawalincluding nicotine, cannabis, amphetamine, cocaine, Attention DeficitHyperactivity Disorder (ADHD) and dyskinetic disorders such asParkinson's disease, neuroleptic-induced parkinsonism, and tardivedyskinesias, schizophrenia, schizophreniform diseases, psychoticdepression, mania, acute mania, paranoid, hallucinogenic and delusionaldisorders, personality disorders, obsessive compulsive disorders,schizotypal disorders, delusional disorders, psychosis due tomalignancy, metabolic disorder, endocrine disease or narcolepsy,psychosis due to drug abuse or drug withdrawal, bipolar disorders,epilepsy and schizo-affective disorder.

2.20 A compound according to any one of Embodiments 1.1 to 1.180 for usein the treatment of Alzheimer's disease.

2.21 A compound according to any one of Embodiments 1.1 to 1.180 for usein the treatment of Schizophrenia.

2.22 A compound according to any one of Embodiments 1.1 to 1.180 for usein the treatment of Alzheimer's disease and/or dementia with Lewybodies.

2.23 A method of treatment of a cognitive disorder in a subject (e.g. amammalian patient such as a human, e.g. a human in need of suchtreatment), which method comprises the administration of atherapeutically effective dose of a compound according to any one ofEmbodiments 1.1 to 1.180.

2.24 A method according to Embodiment 2.20 wherein the cognitivedisorder comprises, arises from or is associated with a condition asdefined in Embodiment 2.19.

2.25 A method according to Embodiment 2.24 wherein the cognitivedisorder arises from or is associated with Alzheimer's disease.

2.26 A method according to Embodiment 2.24 wherein the cognitivedisorder is Schizophrenia.

2.27 The use of a compound according to any one of Embodiments 1.1 to1.180 for the manufacture of a medicament for the treatment of acognitive disorder.

2.28 The use according to Embodiment 2.27 wherein the cognitive disordercomprises, arises from or is associated with a condition as defined inEmbodiment 2.11.

2.29 The use according to Embodiment 2.28 wherein the cognitive disorderarises from or is associated with Alzheimer's disease.

2.30 The use according to Embodiment 2.28 wherein the cognitive disorderis Schizophrenia.

2.31 A compound according to any one of Embodiments 1.1 to 1.180 for thetreatment or lessening the severity of acute, chronic, neuropathic, orinflammatory pain, arthritis, migraine, cluster headaches, trigeminalneuralgia, herpetic neuralgia, general neuralgias, visceral pain,osteoarthritis pain, postherpetic neuralgia, diabetic neuropathy,radicular pain, sciatica, back pain, head or neck pain, severe orintractable pain, nociceptive pain, breakthrough pain, postsurgicalpain, or cancer pain.

2.32 A method of treatment or lessening the severity of acute, chronic,neuropathic, or inflammatory pain, arthritis, migraine, clusterheadaches, trigeminal neuralgia, herpetic neuralgia, general neuralgias,visceral pain, osteoarthritis pain, postherpetic neuralgia, diabeticneuropathy, radicular pain, sciatica, back pain, head or neck pain,severe or intractable pain, nociceptive pain, breakthrough pain,postsurgical pain, or cancer pain, which method comprises theadministration of a therapeutically effective dose of a compoundaccording to any one of Embodiments 1.1 to 1.180.

2.33 A compound according to any one of Embodiments 1.1 to 1.180 for thetreatment of peripheral disorders such as reduction of intra ocularpressure in Glaucoma and treatment of dry eyes and dry mouth includingSjogren's Syndrome.

2.34 A method of treatment of peripheral disorders such as reduction ofintra ocular pressure in Glaucoma and treatment of dry eyes and drymouth including Sjogren's Syndrome, which method comprises theadministration of a therapeutically effective dose of a compoundaccording to any one of Embodiments 1.1 to 1.180.

2.35 The use of a compound according to any one of Embodiments 1.1 to1.180 for the manufacture of a medicament for the treatment or lesseningthe severity of acute, chronic, neuropathic, or inflammatory pain,arthritis, migraine, cluster headaches, trigeminal neuralgia, herpeticneuralgia, general neuralgias, visceral pain, osteoarthritis pain,postherpetic neuralgia, diabetic neuropathy, radicular pain, sciatica,back pain, head or neck pain, severe or intractable pain, nociceptivepain, breakthrough pain, postsurgical pain, or cancer pain or for thetreatment of peripheral disorders such as reduction of intra ocularpressure in Glaucoma and treatment of dry eyes and dry mouth includingSjogren's Syndrome.

2.36 The use of a compound according to any one of Embodiments 1.1 to1.180 for the treatment of addicition.

2.37 The use of a compound according to any one of Embodiments 1.1 to1.180 for the treatment of movement disorders such as Parkinson'sdisease, ADHD, Huntingdon's disease, tourette's syndrome and othersyndromes associated with dopaminergic dysfunction as an underlyingpathogenetic factor driving disease.

Methods for the Preparation of Compounds of the Formula (1) and Formula(1a)

Compounds of the formula (1) and formula (1a) can be prepared inaccordance with synthetic methods well known to the skilled person andas described herein.

Accordingly, in another embodiment (Embodiment 3.1), the inventionprovides a process for the preparation of a compound as defined in anyone of Embodiments 1.1 to 1.180, which process comprises:

(A) the reaction of a compound of the formula (10)

with a compound of the formula (11) or (11a):

under reductive amination conditions; wherein p, q, r, s, R³, R⁴ and Qare as defined in any one of Embodiments 1.1 to 1.180; or

(B) the reaction of a compound of the formula (12) or (12a):

with a compound of the formula Cl—C(═O)—CH₂—R⁴, in the presence of abase; or

(C) the reaction of a compound of the formula (10)

with a compound of the formula (13) or (13a):

under nucleophilic substitution conditions; wherein p, q, r, s, R³, R⁴and Q are as defined in any one of Embodiments 1.1 to 1.180; andoptionally:

(D) converting one compound of the formula (1) or formula (1a) toanother compound of the formula (1) or formula (1a) respectively.

In process variant (A), the piperidine heterocycle (10) is reacted withthe substituted ketone (11) or (11a) under reductive aminationconditions. The reductive amination reaction is typically carried out atambient temperature using a borohydride reducing agent such as sodiumtriacetoxy-borohydride in a solvent such as dichloromethane ordichloroethane containing acetic acid.

In process variant (C), the piperidine heterocycle (10) is reacted withthe sulfonic ester (13 or 13a, R=methyl, trifluoromethyl or4-methylphenyl) in a nucleophilic substitution reaction which istypically carried out with mild heating (e.g. to a temperature of fromabout 40° C. to about 70° C.) either neat, with no solvent, or in asuitable solvent such as tetrahydrofuran, acetonitrile ordimethylacetamide.

Intermediate compounds of the formula (12) and (12a) can be prepared bythe series of reactions shown in Scheme 1 and Scheme 1a respectivelybelow.

In reaction Scheme 1 or Scheme 1a, the piperidine heterocycle (10) isreacted with the Boc-protected spiroketone (14) or (14a) respectivelyunder reductive amination conditions. The reductive amination reactionis typically carried out with mild heating (e.g. to a temperature offrom about 40° C. to about 70° C.) in the presence of either sodiumcyanoborohydride in combination with zinc chloride or sodiumtriacetoxyborohydride in combination with titanium isopropoxide in asolvent such as dichloromethane or dichloroethane containing acetic acidto give an intermediate piperidine compound (15) or (15a) which is thendeprotected by removal of the Boc group by treatment with acid (e.g.trifluoroacetic acid in dichloromethane) to give the compound (12) or(12a) respectively.

Compounds of the formula (12) and (12a) can also be prepared by thesequence of reactions shown in Scheme 2 and Scheme 2a respectivelybelow.

In Scheme 2 or Scheme 2a, the Boc-protected spiroketone (14) or (14a)respectively is reduced to the alcohol (16) or (16a) respectively usingsodium borohydride in methanol. The alcohol (16) or (16a) is thenactivated as the sulfonic ester (17 or 17a respectively, R=methyl,trifluoromethyl or 4-methylphenyl) using the corresponding sulfonylchloride in dichloromethane in the presence of a tertiary amine such astriethylamine or N,N-diisopropylethylamine. The sulfonic ester (17) or(17a) is reacted with the piperidine heterocycle (10) in a nucleophilicsubstitution reaction which is typically carried out with mild heating(e.g. to a temperature of from about 40° C. to about 70° C.) eitherneat, with no solvent, or in a suitable solvent such as tetrahydrofuran,acetonitrile or dimethylacetamide to give compound (15) or (15a)respectively which is then deprotected by removal of the Boc group bytreatment with acid (e.g. trifluoroacetic acid in dichloromethane) togive the compound (12) or (12a) respectively.

Once formed, one compound of the formula (1) or formula (1a), or aprotected derivative thereof, can be converted into another compound ofthe formula (1) or formula (1a) respectively by methods well known tothe skilled person. Examples of synthetic procedures for converting onefunctional group into another functional group are set out in standardtexts such as Advanced Organic Chemistry and Organic Syntheses (seereferences above) or Fiesers' Reagents for Organic Synthesis, Volumes1-17, John Wiley, edited by Mary Fieser (ISBN: 0-471-58283-2). Examplesof these transformations include amide bond formation, urea formation,carbamate formation, alkylation reactions, N-arylation reaction and C—Cbond coupling reactions.

In many of the reactions described above, it may be necessary to protectone or more groups to prevent reaction from taking place at anundesirable location on the molecule. Examples of protecting groups, andmethods of protecting and deprotecting functional groups, can be foundin Protective Groups in Organic Synthesis (T. Greene and P. Wuts; 3rdEdition; John Wiley and Sons, 1999).

Compounds made by the foregoing methods may be isolated and purified byany of a variety of methods well known to those skilled in the art andexamples of such methods include recrystallisation and chromatographictechniques such as column chromatography (e.g. flash chromatography) andHPLC.

Pharmaceutical Formulations

While it is possible for the active compound to be administered alone,it is preferable to present it as a pharmaceutical composition (e.g.formulation).

Accordingly, in another embodiment (Embodiment 4.1) of the invention,there is provided a pharmaceutical composition comprising at least onecompound of the formula (1) or formula (1a) as defined in any one ofEmbodiments 1.1 to 1.180 together with at least one pharmaceuticallyacceptable excipient.

In one embodiment (Embodiment 4.2), the composition is a tabletcomposition.

In another embodiment (Embodiment 4.3), the composition is a capsulecomposition.

The pharmaceutically acceptable excipient(s) can be selected from, forexample, carriers (e.g. a solid, liquid or semi-solid carrier),adjuvants, diluents (e.g solid diluents such as fillers or bulkingagents; and liquid diluents such as solvents and co-solvents),granulating agents, binders, flow aids, coating agents,release-controlling agents (e.g. release retarding or delaying polymersor waxes), binding agents, disintegrants, buffering agents, lubricants,preservatives, anti-fungal and antibacterial agents, antioxidants,buffering agents, tonicity-adjusting agents, thickening agents,flavouring agents, sweeteners, pigments, plasticizers, taste maskingagents, stabilisers or any other excipients conventionally used inpharmaceutical compositions.

The term “pharmaceutically acceptable” as used herein means compounds,materials, compositions, and/or dosage forms which are, within the scopeof sound medical judgment, suitable for use in contact with the tissuesof a subject (e.g. a human subject) without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio. Each excipient mustalso be “acceptable” in the sense of being compatible with the otheringredients of the formulation.

Pharmaceutical compositions containing compounds of the formula (1) orformula (1a) can be formulated in accordance with known techniques, seefor example, Remington's Pharmaceutical Sciences, Mack PublishingCompany, Easton, Pa., USA.

The pharmaceutical compositions can be in any form suitable for oral,parenteral, topical, intranasal, intrabronchial, sublingual, ophthalmic,otic, rectal, intra-vaginal, or transdermal administration.

Pharmaceutical dosage forms suitable for oral administration includetablets (coated or uncoated), capsules (hard or soft shell), caplets,pills, lozenges, syrups, solutions, powders, granules, elixirs andsuspensions, sublingual tablets, wafers or patches such as buccalpatches.

Tablet compositions can contain a unit dosage of active compoundtogether with an inert diluent or carrier such as a sugar or sugaralcohol, eg; lactose, sucrose, sorbitol or mannitol; and/or a non-sugarderived diluent such as sodium carbonate, calcium phosphate, calciumcarbonate, or a cellulose or derivative thereof such as microcrystallinecellulose (MCC), methyl cellulose, ethyl cellulose, hydroxypropyl methylcellulose, and starches such as corn starch. Tablets may also containsuch standard ingredients as binding and granulating agents such aspolyvinylpyrrolidone, disintegrants (e.g. swellable crosslinked polymerssuch as crosslinked carboxymethylcellulose), lubricating agents (e.g.stearates), preservatives (e.g. parabens), antioxidants (e.g. BHT),buffering agents (for example phosphate or citrate buffers), andeffervescent agents such as citrate/bicarbonate mixtures. Suchexcipients are well known and do not need to be discussed in detailhere.

Tablets may be designed to release the drug either upon contact withstomach fluids (immediate release tablets) or to release in a controlledmanner (controlled release tablets) over a prolonged period of time orwith a specific region of the GI tract.

The pharmaceutical compositions typically comprise from approximately 1%(w/w) to approximately 95%, preferably % (w/w) active ingredient andfrom 99% (w/w) to 5% (w/w) of a pharmaceutically acceptable excipient(for example as defined above) or combination of such excipients.Preferably, the compositions comprise from approximately 20% (w/w) toapproximately 90% (w/w) active ingredient and from 80% (w/w) to 10% of apharmaceutically excipient or combination of excipients. Thepharmaceutical compositions comprise from approximately 1% toapproximately 95%, preferably from approximately 20% to approximately90%, active ingredient. Pharmaceutical compositions according to theinvention may be, for example, in unit dose form, such as in the form ofampoules, vials, suppositories, pre-filled syringes, dragées, powders,tablets or capsules.

Tablets and capsules may contain, for example, 0-20% disintegrants, 0-5%lubricants, 0-5% flow aids and/or 0-99% (w/w) fillers/or bulking agents(depending on drug dose). They may also contain 0-10% (w/w) polymerbinders, 0-5% (w/w) antioxidants, 0-5% (w/w) pigments. Slow releasetablets would in addition typically contain 0-99% (w/w)release-controlling (e.g. delaying) polymers (depending on dose). Thefilm coats of the tablet or capsule typically contain 0-10% (w/w)polymers, 0-3% (w/w) pigments, and/or 0-2% (w/w) plasticizers.

Parenteral formulations typically contain 0-20% (w/w) buffers, 0-50%(w/w) cosolvents, and/or 0-99% (w/w) Water for Injection (WFI)(depending on dose and if freeze dried). Formulations for intramusculardepots may also contain 0-99% (w/w) oils.

The pharmaceutical formulations may be presented to a patient in“patient packs” containing an entire course of treatment in a singlepackage, usually a blister pack.

The compounds of the formula (1) or formula (1a) will generally bepresented in unit dosage form and, as such, will typically containsufficient compound to provide a desired level of biological activity.For example, a formulation may contain from 1 nanogram to 2 grams ofactive ingredient, e.g. from 1 nanogram to 2 milligrams of activeingredient. Within these ranges, particular sub-ranges of compound are0.1 milligrams to 2 grams of active ingredient (more usually from 10milligrams to 1 gram, e.g. 50 milligrams to 500 milligrams), or 1microgram to 20 milligrams (for example 1 microgram to 10 milligrams,e.g. 0.1 milligrams to 2 milligrams of active ingredient).

For oral compositions, a unit dosage form may contain from 1 milligramto 2 grams, more typically 10 milligrams to 1 gram, for example 50milligrams to 1 gram, e.g. 100 miligrams to 1 gram, of active compound.

The active compound will be administered to a patient in need thereof(for example a human or animal patient) in an amount sufficient toachieve the desired therapeutic effect (effective amount). The preciseamounts of compound administered may be determined by a supervisingphysician in accordance with standard procedures.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows the locomotor activity observed in the hyperactivity testof Example B.

EXAMPLES

The invention will now be illustrated, but not limited, by reference tothe specific embodiments described in the following examples.

Examples 1-1 to 3-3

The compounds of Examples 1-1 to 3-3 shown in Table 1 below have beenprepared. Their NMR and LCMS properties and the methods used to preparethem are set out in Table 3.

TABLE 1

Example 1-1

Example 1-2

Example 2-1

Example 2-2

Example 2-3

Example 2-4

Example 2-5

Example 2-6

Example 2-7

Example 2-8

Example 2-9

Example 2-10

Example 2-11

Example 2-12

Example 2-13

Example 2-14

Example 2-15

Example 2-16

Example 2-17

Example 2-18

Example 2-19

Example 2-20

Example 2-21

Example 2-22

Example 2-23

Example 2-24

Example 2-25

Example 2-26

Example 2-27

Example 2-28

Example 2-29

Example 2-30

Example 2-31

Example 2-32

Example 2-33

Example 2-34

Example 2-35

Example 2-36

Example 2-37

Example 2-38

Example 2-39

Example 2-40

Example 2-41

Example 2-42

Example 2-43

Example 2-44

Example 2-45

Example 2-46

Example 2-47

Example 2-48

Example 2-49

Example 2-50

Example 2-51

Example 2-52

Example 2-53

Example 2-54

Example 2-55

Example 2-56

Example 2-57

Example 2-58

Example 2-59

Example 2-60

Example 2-61

Example 2-62

Example 2-63

Example 2-64

Example 2-65

Example 2-66

Example 2-67

Example 3-1

Example 3-2

Example 3-3

General Procedures

Where no preparative routes are included, the relevant intermediate iscommercially available. Commercial reagents were utilized withoutfurther purification. Room temperature (rt) refers to approximately20-27° C. ¹H NMR spectra were recorded at 400 MHz on either a Bruker orJeol instrument. Chemical shift values are expressed in parts permillion (ppm), i.e. (δ:)-values. The following abbreviations are usedfor the multiplicity of the NMR signals: s=singlet, br=broad, d=doublet,t=triplet, q=quartet, quint=quintet, td=triplet of doublets, tt=tripletof triplets, qd=quartet of doublets, ddd=doublet of doublet of doublets,ddt=doublet of doublet of triplets, m=multiplet. Coupling constants arelisted as Jvalues, measured in Hz. NMR and mass spectroscopy resultswere corrected to account for background peaks. Chromatography refers tocolumn chromatography performed using 60-120 mesh silica gel andexecuted under nitrogen pressure (flash chromatography) conditions. TLCfor monitoring reactions refers to TLC run using the specified mobilephase and Silica gel F254 (Merck) as a stationary phase.Microwave-mediated reactions were performed in Biotage Initiator or CEMDiscover microwave reactors.

LCMS experiments were typically carried out using electrosprayconditions as specified for each compound under the followingconditions:

LCMS Method C

Instruments: Agilent 1260 Infinity LC with Diode Array Detector, Agilent6120B Single Quadrupole MS with API-ES Source; Column: PhenomenexGemini-NX C-18, 3 micron, 2.0×30 mm; Gradient [time (min)/solvent B in A(%)]: Method: 0.00/5, 2.00/95, 2.50/95, 2.60/5, 3.00/5; Solvents:solvent A=2.5 L H₂O+2.5 mL of (28% NH₃ in H₂O); solvent B=2.5 L MeCN+129mL H₂O+2.7 mL of (28% NH₃ in H₂O); Injection volume 0.5 μL; UV detection190 to 400 nM; column temperature 40° C.; Flow rate 1.5 mL/min.

LCMS Methods D and E

Instruments: HP 1100 with G1315A DAD, Micromass ZQ; Column: WatersX-Bridge C-18, 2.5 micron, 2.1×20 mm or Phenomenex Gemini-NX C-18, 3micron, 2.0×30 mm; Gradient [time (min)/solvent D in C (%)]: Method D:0.00/2, 0.10/2, 2.50/95, 3.50/95 or Method E: 0.00/2, 0.10/2, 8.40/95,10.00/95; Solvents: solvent C=2.5 L H₂O+2.5 mL 28% ammonia in H₂Osolution; solvent D=2.5 L MeCN+135 mL H₂O+2.5 mL 28% ammonia in H₂Osolution); Injection volume 1 μL; UV detection 230 to 400 nM; Massdetection 130 to 800 AMU (+ve and −ve electrospray); column temperature45° C.; Flow rate 1.5 mL/min.

LCMS Method F

Instruments: Waters Acquity H Class, Photo Diode Array, SQ Detector;Column: BEH C18, 1.7 micron, 2.1×50 mm; Gradient [time (min)/solvent Bin A (%)]: 0.00/5, 0.40/5, 0.8/35, 1.20/55, 2.50/100, 3.30/100 4.00/5;Solvents: solvent A=5 mM mmmonium acetate and 0.1% formic acid in H₂O;solvent B=0.1% formic acid in MeCN; Injection volume 2 μL; UV detection200 to 400 nM; Mass detection 100 to 1200 AMU (+ve electrospray); columnat ambient temperature; Flow rate 0.5 mL/min.

LCMS Method H

Instruments: Waters 2695, Photo Diode Array, ZQ-2000 Detector; Column:X-Bridge C18, 5 micron, 150×4.6 mm; Gradient [time (min)/solvent B in A(%)]: 0.00/100, 7.00/50, 9.00/0, 11.00/0, 11.01/100, 12.00/100;Solvents: solvent A=0.1% ammonia in H₂O; solvent B=0.1% ammonia in MeCN;Injection volume 10 μL; UV detection 200 to 400 nM; Mass detection 60 to1000 AMU (+ve electrospray); column at ambient temperature; Flow rate1.0 mL/min.

LCMS Method I

Instruments: Waters 2695, Photo Diode Array, ZQ-2000 Detector; Column:X-Bridge C18, 3.5 micron, 150×4.6 mm; Gradient [time (min)/solvent B inA (%)]: 0.00/5, 5.00/90, 5.80/95, 10/95; Solvents: solvent A=0.1%ammonia in H₂O; solvent B=0.1% ammonia in MeCN; Injection volume 10 μL;UV detection 200 to 400 nM; Mass detection 60 to 1000 AMU (+veelectrospray); column at ambient temperature; Flow rate 1.0 mL/min.

LCMS Method K

Instruments: Waters 2695, Photo Diode Array, ZQ-2000 Detector; Column:X-Bridge C18, 3.5 micron, 50×4.6 mm; Gradient [time (min)/solvent B in A(%)]: 0.01/0, 0.20/0, 5.00/90, 5.80/95, 7.20/95, 7.21/100, 10.00/100;Solvents: solvent A=0.1% ammonia in H₂O; solvent B=0.1% ammonia in MeCN;Injection volume 10 μL; UV detection 200 to 400 nM; Mass detection 60 to1000 AMU (+ve electrospray); column at ambient temperature; Flow rate1.0 mL/min.

LCMS data in the experimental section are given in the format: Mass ion,retention time, UV activity.

Abbreviations

-   AcOH=acetic acid-   CDI=1,1′-Carbonyldiimidazole-   d=day(s)-   DAST=diethylaminosulfur trifluoride-   DCE=dichloroethane-   DCM=dichloromethane-   DIPEA=diisopropylethylamine-   DIAD=diisopropyl azodicarboxylate-   DMF=dimethylformamide-   DMP=Dess-Martin periodinane-   DMSO=dimethylsulfoxide-   ES=electro spray ionisation-   EtOAc=ethyl acetate-   h=hour(s)-   HATU=1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium    3-oxid hexafluorophosphate-   HPLC=high performance liquid chromatography-   LC=liquid chromatography-   LiAlH₄/LAH=Lithium aluminium hydride-   MeCN=acetonitrile-   MeOH=methanol-   min=minute(s)-   MS=mass spectrometry-   Et₃N=triethylamine-   NMR=nuclear magnetic resonance-   rt=room temperature-   sat.=saturated-   sol.=solution-   STAB=sodium triacetoxyborohydride-   THF=tetrahydrofuran-   TLC=thin layer chromatography

Prefixes n-, s-, i-, t- and tert- have their usual meanings: normal,secondary, iso, and tertiary.

Final compounds are named using the software package ACD/ChemSketchVersion 12. Intermediates and reagents are named either using thesoftware package ACD/ChemSketch Version 12 or are referred to usingtheir common name as typically found in suppliers catalogues etc.

General Synthetic Procedures for the Intermediates

Route 1

Procedure for the Preparation of Intermediate 4, ethyl2-oxo-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 3, tert-butyl 2-oxo-6-azaspiro[3.4]octane-6-carboxylate(3.37 g, 15 mmol) was added portionwise to hydrogen chloride (4 Mdioxane solution, 50 mL, 200 mmol). Caution: effervescence. After 24 h,the reaction was concentrated in vacuo and the residual solid wasdissolved in a mixture of Et₃N (4.18 mL, 30 mmol) and DCM (66 mL). Oncompletion of dissolution, the solution was immediately cooled to 0° C.,then Intermediate 5, ethyl carbonochloridate (1.57 mL, 16 mmol) wasadded dropwise. After 18 h, the mixture was poured into DCM (100 mL) andNaHCO₃ (aq) (100 mL) and extracted with DCM (2×100 mL). The organiclayers were collected, washed with brine (20 mL), dried over MgSO₄, thenthe residue after evaporation was purified by column chromatography(normal phase, [Biotage SNAP cartridge KP-sil 100 g, 40-63 μm, 60 Å, 50mL per min, gradient 0% to 4% MeOH in DCM]) to give Intermediate 4,ethyl 2-oxo-6-azaspiro[3.4]octane-6-carboxylate as an oil (2.47 g, 83%).The data for the title compound are in Table 2.

Route 2

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 2, N-ethyl-N-(piperidin-4-yl)acetamidehydrochloride

To Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (3.0 g, 15.1mmol) in MeOH (40 mL) was added Intermediate 44, ethanamine (12.6 mL,25.1 mmol, 2 M in THF), Et₃N (4.2 mL, 30.3 mmol) and ZnCl₂ (0.1 g, 0.7mmol) and the reaction mixture was stirred at 60° C. for 7 h. NaBH₃CN(1.2 g, 19.6 mmol) was added portionwise and the resulting reactionmixture was stirred at 25° C. for 17 h. The solvents were removed invacuo and the residue was partitioned between H₂O (250 mL) and EtOAc(200 mL). The aqueous layer was further extracted with EtOAc (2×200 mL),and the combined organic phases were dried (Na₂SO₄) and the solvent wasremoved in vacuo. The residue was purified by column chromatography(normal basic activated alumina, 10 to 30% EtOAc in hexane) to giveIntermediate 43, tert-butyl 4-(ethylamino)piperidine-1-carboxylate (3.0g, 88%) as a gum.

The data for Intermediate 43 are in Table 2.

To Intermediate 43, tert-butyl 4-(ethylamino)piperidine-1-carboxylate(0.20 g, 0.9 mmol) in DCM (10 mL) was added triethylamine (0.15 mL, 1.1mmol) dropwise and the mixture was stirred at 0° C. for 30 min. Acetylchloride (0.09 g, 1.1 mmol) was added dropwise at 0° C. and theresulting reaction mixture was stirred at 25° C. for 8 h before removalof the solvents in vacuo. The residue was partitioned between H₂O (120mL) and EtOAc (100 mL) and the aqueous layer was further extracted withEtOAc (2×100 mL). The combined organic phases were dried (Na₂SO₄) andthe solvent removed in vacuo. The residue was purified by columnchromatography (normal basic activated alumina, 0.5 to 1.0% MeOH in DCM)to give tert-butyl 4-[acetyl(ethyl)amino]piperidine-1-carboxylate (0.15g, 63%) as a liquid.

LCMS (Method I): m/z 271 [M+H]⁺ (ES⁺), at 3.79 min, UV active.

To tert-butyl 4-[acetyl(ethyl)amino]piperidine-1-carboxylate (0.20 g,0.7 mmol) in 1,4-dioxane (5 mL) was added 4.0 M HCl in 1,4-dioxane (5mL) dropwise and the resulting reaction mixture was stirred at 25° C.for 16 h. The solvent was removed in vacuo and the residue was purifiedby triturating with diethyl ether (3×5 mL) to give Intermediate 2,N-ethyl-N-(piperidin-4-yl)acetamide hydrochloride salt (0.15 g, 100%) asa solid.

The data for the title compound is in Table 2.

Route 3

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 8, N-cyclopropyl-N-(piperidin-4-yl)acetamide

Intermediate 7, cyclopropanamine (1.2 g, 21.5 mmol), Intermediate 6,benzyl 4-oxopiperidine-1-carboxylate (5.0 g, 21.5 mmol), and zincchloride (0.15 g, 1.1 mmol) were dissolved in MeOH (15 mL) and heated to50-60° C. for 3 h under N₂. The mixture was then cooled to 0-10° C.before portionwise addition of NaCNBH₃ (1.8 g, 27.9 mmol) and furtherstirring at rt for 2 h. The reaction mixture was partitioned between H₂O(15 mL) and EtOAc (25 mL) and the aqueous layer was further extractedwith EtOAc (2×25 mL). The combined organic phases were dried (Na₂SO₄)and the solvent was removed in vacuo to yield benzyl4-(cyclopropylamino)piperidine-1-carboxylate (4.0 g, 68%) as a gum.

LCMS (Method F): m/z 275 [M+H]⁺ (ES⁺), at 1.60 min, UV active

To benzyl 4-(cyclopropylamino)piperidine-1-carboxylate (2.5 g, 9.1 mmol)in DCM (10 mL) cooled to 0-5° C. was added Et₃N (1.8 g, 18.2 mmol) andacetyl chloride (0.9 g, 11.9 mmol) dropwise and the reaction mixture wasstirred at 25° C. for 16 h. The reaction mixture was partitioned betweenH₂O (15 mL) and DCM (25 mL) and the aqueous layer was further extractedwith DCM (2×25 mL). The combined organic phases were dried (Na₂SO₄) andthe solvent was removed in vacuo to yield benzyl4-[acetyl(cyclopropyl)amino]piperidine-1-carboxylate (2.0 g, 70%) as agum, which was used in the next step without further purification.

LCMS (Method I): m/z 317 [M+H]⁺ (ES⁺), at 4.17 min, UV active

To benzyl 4-[acetyl(cyclopropyl)amino]piperidine-1-carboxylate (2.0 g,6.3 mmol) in MeOH (15 mL) was added 10% Pd/C (0.2 g) at rt and thereaction mixture was stirred under an atmosphere H₂ gas (10 kg pressure)for 16 h at rt. The reaction mixture was then filtered through Celiteand the solvent was removed from the filtrate in vacuo to yieldIntermediate 8, N-cyclopropyl-N-(piperidin-4-yl) acetamide (1.0 g, 86%)as a gum, which was used in the next step without further purification.

The data for the title compound are in Table 2.

Route 4

Typical Procedure for the Preparation of Amines Substituted with aPendant Ester, as Exemplified by the Preparation of Intermediate 13,methyl [cyclopropyl(piperidin-4-yl)amino]acetate hydrochloride

To Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (5.0 g, 25.1mmol) in MeOH (40 mL) was added Intermediate 7, cyclopropanamine (1.4 g,25.1 mmol), Et₃N (10.0 mL, 75.3 mmol) and ZnCl₂ (0.3 g, 2.5 mmol). Thereaction mixture was stirred at 60° C. for 7 h, then NaBH₃CN (4.8 g,75.3 mmol) was added portionwise. The resulting reaction mixture wasstirred at 25° C. for 17 h. The solvents were removed in vacuo, and theresidue was partitioned between H₂O (250 mL) and EtOAc (200 mL). Theaqueous layer was extracted further with EtOAc (2×200 mL), the combinedorganic layers were dried (Na₂SO₄) and the solvent was removed in vacuo.The residue was purified by column chromatography (normal basicactivated alumina, 10% to 30% EtOAc in hexane) to give Intermediate 11,tert-butyl 4-(cyclopropylamino)piperidine-1-carboxylate (5.3 g, 88%) asa gum.

The data for Intermediate 11 are in Table 2.

Intermediate 11, tert-butyl 4-(cyclopropylamino)piperidine-1-carboxylate(300 mg, 1.25 mmol) was dissolved in DMF (10 mL) and K₂CO₃ (517 mg, 3.75mmol) was added. The reaction mixture was stirred at 70° C. for 3 h,then Intermediate 12, methyl bromoacetate (229 mg, 1.50 mmol) was addeddropwise at 20° C. The resulting reaction mixture was stirred at 60° C.for 8 h. The solvents were removed in vacuo and the residue waspartitioned between H₂O (150 mL) and EtOAc (100 mL). The aqueous layerwas further extracted with EtOAc (2×100 mL), and the combined organiclayers were dried (Na₂SO₄) and the solvents were removed in vacuo. Theresidue was purified by column chromatography (normal basic activatedalumina, 0.5% to 1.0% MeOH in DCM) to give tert-butyl4-[cyclopropyl(2-methoxy-2-oxoethyl)amino]piperidine-1-carboxylate (310mg, 80%) as a gum.

LCMS (Method I): m/z 313 [M+H]⁺ (ES⁺), at 4.85 min, UV active

tert-Butyl4-[cyclopropyl(2-methoxy-2-oxoethyl)amino]piperidine-1-carboxylate (300mg, 0.96 mmol) was dissolved in 1,4-dioxane (5 mL) and 4 M HCl in1,4-dioxane (3 mL) was added dropwise. The resulting reaction mixturewas stirred at 25° C. for 16 h. The solvents were removed in vacuo, andthe residue was purified by triturating with diethyl ether (3×5 mL) togive Intermediate 13, methyl [cyclopropyl(piperidin-4-yl)amino]acetatehydrochloride salt (210 mg, 85%) as a solid.

The data for the title compound are in Table 2.

Route 5

Typical Procedure for the Preparation of Amines Substituted with anArylmethyl Group, as Exemplified by the Preparation of Intermediate 15,N-cyclopropyl-N-(1,2-oxazol-3-ylmethyl)piperidin-4-amine hydrochloride

Intermediate 11, tert-butyl 4-(cyclopropylamino)piperidine-1-carboxylate(200 mg, 0.83 mmol) was dissolved in ethanol (10 mL) and sodiumbicarbonate (200 mg, 2.38 mmol) was added. The reaction mixture wasstirred at 0° C. for 30 min, then Intermediate 14,3-(chloromethyl)-1,2-oxazole (97 mg, 0.83 mmol) was added dropwise atrt. The resulting reaction mixture was stirred at 60° C. for 16 h. Thesolvents were removed in vacuo and the residue was partitioned betweenH₂O (120 mL) and EtOAc (100 mL). The aqueous layer was further extractedwith EtOAc (2×100 mL) and the combined organic layers were dried(Na₂SO₄), and the solvents were removed in vacuo. The residue waspurified by column chromatography (normal basic activated alumina, 0.5%to 1.0% MeOH in DCM) to give tert-butyl4-[cyclopropyl(1,2-oxazol-3-ylmethyl)amino]piperidine-1-carboxylate (150mg, 58%) as a liquid.

LCMS (Method I): m/z 322 [M+H]⁺ (ES⁺), at 4.92 min, UV active

tert-Butyl4-[cyclopropyl(1,2-oxazol-3-ylmethyl)amino]piperidine-1-carboxylate (150mg, 0.46 mmol) was dissolved in 1,4-dioxane (5 mL) and 4 M HCl in1,4-dioxane (5 mL) was added dropwise. The resulting reaction mixturewas stirred at 25° C. for 16 h. The solvents were removed in vacuo, andthe residue was purified by triturating with diethyl ether (3×5 mL) togive Intermediate 15,N-cyclopropyl-N-(1,2-oxazol-3-ylmethyl)piperidin-4-amine hydrochloridesalt (120 mg, 100%) as a solid.

The data for the title compound are in Table 2.

Route 6

Typical Procedure for the Preparation of Amines Substituted with TwoAlkyl Groups, as Exemplified by the Preparation of Intermediate 18,N-ethyl-N-(2,2,2-trifluoroethyl)piperidin-4-amine hydrochloride

To Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (500 mg, 2.5mmol) as a solution in MeOH (10 mL) was added Intermediate 19,2,2,2-trifluoroethanamine (273 mg, 2.8 mmol), triethylamine (1.0 mL, 7.5mmol) and ZnCl₂ (34 mg, 0.3 mmol) and the reaction mixture was stirredat 60° C. for 7 h. NaBH₃CN (475 mg, 7.5 mmol) was then added portionwiseand the resulting reaction mixture was stirred at 25° C. for 17 h. Thesolvent was removed in vacuo and the residue was partitioned between H₂O(150 mL) and EtOAc (120 mL). The aqueous layer was further extractedwith EtOAc (2×120 mL) and the combined organic phases were dried(Na₂SO₄) and the solvent was removed in vacuo. The residue was purifiedby column chromatography (normal basic activated alumina, 0.5% to 1.0%MeOH in DCM) to give Intermediate 97, tert-butyl4-[(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (350 mg, 49%) asa gum.

The data for Intermediate 97 are in Table 2.

To Intermediate 97, tert-butyl4-[(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (300 mg, 1.1mmol) as a solution in MeOH (10 mL) was added Intermediate 20,acetaldehyde (69 mg, 1.6 mmol), triethylamine (0.4 mL, 3.2 mmol) andZnCl₂ (14 mg, 0.1 mmol) and the reaction mixture was stirred at 50° C.for 7 h. The mixture was allowed to cool to rt before addition ofNaBH₃CN (201 mg, 3.2 mmol) portionwise. The mixture was stirred at 25°C. for 17 h, then the solvent was removed in vacuo. The residue waspartitioned between H₂O (150 mL) and EtOAc (120 mL) and the aqueouslayer was further extracted with EtOAc (2×120 mL). The combined organicphases were dried (Na₂SO₄) and the solvent was removed in vacuo. Theresidue was purified by column chromatography (normal basic activatedalumina, at 0.5% to 3% MeOH in DCM) to give tert-butyl4-[ethyl(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (280 mg,85%) as a gum.

LCMS (Method I): m/z 311 [M+H]⁺ (ES⁺), at 5.65 min, UV active

To tert-butyl4-[ethyl(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (220 mg,0.7 mmol) in 1,4-dioxane (5 mL) was added 4.0 M HCl in 1,4-dioxane (5mL) dropwise and the resulting mixture was stirred at 25° C. for 16 h.The solvents were removed in vacuo and the residue was purified bytrituration with ether (3×5 mL) to give Intermediate 18,N-ethyl-N-(2,2,2-trifluoroethyl)piperidin-4-amine hydrochloride salt(164 mg, 94%) as a solid.

The data for the title compound are in Table 2.

Route 7

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 21,N-cyclopropyl-N-(2-methoxyethyl)piperidin-4-amine hydrochloride

To a solution of Intermediate 11, tert-butyl4-(cyclopropylamino)piperidine-1-carboxylate (0.50 g, 2.1 mmol) inacetonitrile (10 mL) was added Cs₂CO₃ (2.03 g, 6.2 mmol) and CuI (20 mg)and the reaction mixture was stirred at 70° C. for 1 h. Intermediate 22,1-bromo-2-methoxyethane (0.43 g, 3.1 mmol) was then added dropwise at25° C. and the mixture was stirred at 75° C. for 70 h. The solvent wasremoved in vacuo and the residue was partitioned between H₂O (150 mL)and EtOAc (120 mL). The aqueous layer was further extracted with EtOAc(2×120 mL) and the combined organic phases were dried (Na₂SO₄) and thesolvent was removed in vacuo.

The residue was purified by column chromatography (normal basicactivated alumina, 0.5% MeOH in DCM) to give tert-butyl4-[cyclopropyl(2-methoxyethyl)amino]piperidine-1-carboxylate (0.27 g,44%) as a gum.

LCMS (Method I): m/z 299 [M+H]⁺ (ES⁺), at 4.81 min, UV active

To a solution of tert-butyl4-[cyclopropyl(2-methoxyethyl)amino]piperidine-1-carboxylate (0.27 g,0.9 mmol) in 1,4-dioxane (5 mL) was added 4.0 M HCl in 1,4-dioxane (5mL) dropwise and the resulting reaction mixture was stirred at 25° C.for 16 h. The solvent was removed in vacuo and the residue was purifiedby triturating with ether (3×5 mL) to give Intermediate 21,N-cyclopropyl-N-(2-methoxyethyl)piperidin-4-amine hydrochloride salt(0.17 g, 81%) as a solid.

The data for the title compound are in Table 2.

Route 8

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 35,N-cyclopropyl-N-(piperidin-4-ylmethyl)acetamide hydrochloride

To Intermediate 34, tert-butyl 4-formylpiperidine-1-carboxylate (427 mg,2.0 mmol) and Intermediate 7, cyclopropanamine (114 mg, 2.0 mmol) as asolution in DCM (10 mL) at rt was added AcOH (0.23 mL, 4.0 mmol). Themixture was stirred for 3 h then STAB (1.06 g, 5.0 mmol) was added andthe mixture was stirred at rt overnight. The reaction mixture wasquenched with the addition of sat. aq. NaHCO₃ (20 mL). Solid Na₂CO₃ wasadded to ensure the aqueous layer was basic, then the reaction mixturewas extracted with DCM (4×20 mL). The organic layers were combined,dried (MgSO₄), filtered and the solvents were removed in vacuo to givecrude tert-butyl 4-[(cyclopropylamino)methyl]piperidine-1-carboxylate(assumed 100%) which was used directly without further purification.

LCMS (Method C): m/z 255 (M+H)⁺ (ES⁺), at 1.38 min, UV active.

To a solution of tert-butyl4-[(cyclopropylamino)methyl]piperidine-1-carboxylate (assumed 2.0 mmol)in DCM (10 mL) was added Et₃N (1.12 mL, 8.0 mmol), HATU (914 mg, 2.4mmol) and AcOH (0.23 mL, 4.0 mmol) and the reaction mixture was stirredovernight. The mixture was quenched with the addition of sat. aq. NaHCO₃(20 mL) and extracted with DCM (4×20 mL). The organic layers werecombined, dried (MgSO₄), filtered and the solvents were removed in vacuoto give crude tert-butyl4-{[acetyl(cyclopropyl)amino]methyl}piperidine-1-carboxylate (assumed100%) which was used directly without further purification.

LCMS (Method C): m/z 319 (M+Na)⁺ (ES⁺), at 1.26 min, UV active.

To a suspension of tert-butyl4-{[acetyl(cyclopropyl)amino]methyl}piperidine-1-carboxylate (assumed2.0 mmol) in DCM (10 mL) was added 4.0 M HCl in 1,4-dioxane (2.5 mL,10.0 mmol) and the mixture was stirred at rt overnight. The solventswere removed in vacuo to give Intermediate 35,N-cyclopropyl-N-(piperidin-4-ylmethyl)acetamide hydrochloride salt(assumed 2.0 mmol) as a solid which was used without furtherpurification.

The data for the title compound are in Table 2

Route 9

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 36,N-cyclopropyl-N-(piperidin-4-ylmethyl)propanamide hydrochloride

To a solution of Intermediate 34, tert-butyl4-formylpiperidine-1-carboxylate (0.43 g, 2.0 mmol) and Intermediate 7,cyclopropanamine (0.11 g, 2.0 mmol) in DCM (10 mL) at rt was added AcOH(0.23 mL, 4.0 mmol) and the resulting mixture was stirred for 3 h. STAB(1.06 g, 5.0 mmol) was added and the mixture was stirred at rtovernight. The reaction mixture was quenched with the addition of sat.aq. NaHCO₃ (20 mL), then solid Na₂CO₃ was added to ensure the aqueouslayer was basic. The mixture was extracted with DCM (4×20 mL) and thecombined organic layers were dried (MgSO₄), filtered and the solventswere removed in vacuo to give crude tert-butyl4-[(cyclopropylamino)methyl]piperidine-1-carboxylate (assumed 100%)which was used directly without further purification.

LCMS (Method C): m/z 255 (M+H)⁺ (ES⁺), at 1.38 min, UV active.

To a solution of tert-butyl4-[(cyclopropylamino)methyl]piperidine-1-carboxylate (assumed 2.0 mmol)in DCM (10 mL), Et₃N (1.12 mL, 8.0 mmol) and Intermediate 29, propanoylchloride (0.26 mL, 3.0 mmol) were added and the reaction mixture wasstirred overnight. The mixture was quenched with the addition of sat.aq. NaHCO₃ (20 mL) and extracted with DCM (4×20 mL). The organic layerswere combined, dried (MgSO₄), filtered and the solvents were removed invacuo to give crude tert-butyl4-{[cyclopropyl(propanoyl)amino]methyl}piperidine-1-carboxylate (assumed100%) which was used directly without further purification.

LCMS (Method C): m/z 333 (M+Na)⁺ (ES⁺), at 1.39 min, UV active.

To a suspension of tert-butyl4-{[cyclopropyl(propanoyl)amino]methyl}piperidine-1-carboxylate (assumed2.0 mmol) in DCM (10 mL), 4.0 M HCl in 1,4-dioxane (2.5 mL, 10.0 mmol)was added and the mixture was stirred at rt overnight. The solvents wereremoved in vacuo to give Intermediate 36,N-cyclopropyl-N-(piperidin-4-ylmethyl)propanamide hydrochloride salt,(assumed 100%) as a solid which was used without further purification.

The data for the title compound are in Table 2

Route 10

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 42, N-ethyl-N-(piperidin-4-yl) formamide

Intermediate 52, benzyl 4-(ethylamino)piperidine-1-carboxylate (500 mg,1.91 mmol) and p-toluenesulphonic acid (10 mg, 0.06 mmol) were dissolvedin triethyl orthoformate (3.3 mL, 19.84 mmol) at rt and the reactionmixture was stirred at 80° C. for 16 h. The reaction mixture wasquenched with 0.1 N HCl (30 mL) and extracted with 10% MeOH in DCM (2×30mL). The organic layers were combined, washed with sat. aqueous NaHCO₃(30 mL) and dried (Na₂SO₄). The solvent was removed in vacuo to givecrude benzyl 4-[ethyl(formyl)amino]piperidine-1-carboxylate (400 mg,100%) as a gum, which was used without further purification.

LCMS (Method F): m/z 291 [M+H]⁺ (ES⁺), at 1.86 min, UV active.

Benzyl 4-[ethyl(formyl)amino]piperidine-1-carboxylate (180 mg, 0.62mmol) was dissolved in MeOH (15 mL) and 10% Pd/C (50% moisture) (100 mg,0.09 mmol) was added at rt under a nitrogen atmosphere. The system waspurged of nitrogen and placed under hydrogen gas and stirred at rt for16 h. The catalyst was removed by filtration and the filtrate wasconcentrated in vacuo to give crude Intermediate 42,N-ethyl-N-piperidin-4-ylformamide (100 mg, 100%) as a gum.

The data for the title compound is in Table 2.

Route 11

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 53, N-ethyl-N-(piperidin-4-yl)acetamidetrifluoroacetate

To Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (3.0 g, 15.1mmol) in MeOH (40 mL) was added Intermediate 44, ethanamine (2 M in THF,12.6 mL, 25.1 mmol), Et₃N (4.2 mL, 30.3 mmol) and ZnCl₂ (0.1 g, 0.7mmol) and the reaction mixture was stirred at 60° C. for 7 h. NaBH₃CN(1.2 g, 19.6 mmol) was added portionwise and the resulting reactionmixture was stirred at rt for 17 h. The solvents were removed in vacuoand the residue was partitioned between H₂O (250 mL) and EtOAc (200 mL)and the aqueous layer was further extracted with EtOAc (2×200 mL). Thecombined organic phases were dried (Na₂SO₄), the solvent was removed invacuo and the residue was purified by column chromatography (normalbasic activated alumina, 10 to 30% EtOAc in hexane) to give Intermediate43, tert-butyl 4-(ethylamino)piperidine-1-carboxylate (3.0 g, 88%) as agum.

The data for Intermediate 43 is in Table 2.

To Intermediate 43, tert-butyl 4-(ethylamino)piperidine-1-carboxylate(0.20 g, 0.9 mmol) in DCM (10 mL) was added triethylamine (0.15 mL, 1.1mmol) dropwise and the resulting mixture was stirred at 0° C. for 30min. Acetyl chloride (0.09 g, 1.1 mmol) was then added dropwise at 0° C.and the resulting reaction mixture was stirred at rt for 8 h. Thesolvents were removed in vacuo and the residue was partitioned betweenH₂O (120 mL) and EtOAc (100 mL). The aqueous layer was further extractedwith EtOAc (2×100 mL) and the combined organic phases were dried(Na₂SO₄) and the solvent was removed in vacuo. The crude residue waspurified by column chromatography (normal basic activated alumina, 0.5to 1.0% MeOH in DCM) to give tert-butyl4-[acetyl(ethyl)amino]piperidine-1-carboxylate (0.15 g, 63%) as aliquid.

LCMS (Method I): m/z 271 [M+H]⁺ (ES⁺), at 3.79 min, UV active.

To tert-butyl 4-[acetyl(ethyl)amino]piperidine-1-carboxylate (450 mg,1.66 mmol) in DCM (15 mL) was added trifluoroacetic acid (1.3 mL, 16.66mmol) dropwise at 0° C. The resulting reaction mixture was stirred at rtfor 16 h. The solvent was removed in vacuo and the residue was purifiedby triturating with diethyl ether (3×5 mL) to give Intermediate 53,N-ethyl-N-(piperidin-4-yl)acetamide trifluoroacetate salt (450 mg, 100%)as a gum.

The data for the title compound is in Table 2.

Route 12

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 76,N-methoxy-N-(piperidin-4-yl)acetamide trifluoroacetate

To Intermediate 75, O-methylhydroxylamine (0.5 g, 6.0 mmol) in MeOH (25mL) was added sodium acetate (0.51 g, 6.2 mmol) and the reaction mixturewas stirred at rt for 5 minutes. Intermediate 1, tert-Butyl4-oxopiperidine-1-carboxylate (1.0 g, 5.0 mmol), AcOH (0.5 g, 8.8 mmol)and NaBH₃CN (0.3 g, 5.0 mmol) were added. The resulting reaction mixturewas stirred at rt for 24 h, then partitioned between H₂O (30 mL) andEtOAc (50 mL). The organic phase was washed with sat. aq. NaHCO₃ (20 mL)and sat. aq. NaCl (20 mL). The organic phase was dried (Na₂SO₄), thesolvent was removed in vacuo and the residue was purified by columnchromatography (normal phase silica, EtOAc and hexanes) to givetert-butyl 4-(methoxyamino)piperidine-1-carboxylate (1 g, 90%) as asolid.

LCMS (Method F): m/z 231 [M+H]⁺ (ES)+, at 2.07 min, UV active.

To a stirred solution of tert-butyl4-(methoxyamino)piperidine-1-carboxylate (120 mg, 0.52 mmol) in DCE (3ml) was added Ac₂O (79 mg, 0.78 mmol) and Et₃N (0.1 mL, 0.78 mmol) at 0°C. The reaction mixture was heated to 50-60° C. for 3 h, thenpartitioned between H₂O (5 mL) and DCM (10 mL). The aqueous layer wasfurther extracted with DCM (2×10 mL), and the combined organic phaseswere washed with sat. aq. NaHCO₃ (30 mL) and sat. aq. NaCl (30 mL), thendried (Na₂SO₄) and the solvent was removed in vacuo to give crudetert-butyl 4-[acetyl(methoxy)amino]piperidine-1-carboxylate (110 mg,77%) which was used in the next step without further purification.

LCMS (Method F): m/z 273 [M+H]⁺ (ES)+, at 2.02 min, UV active

To a stirred solution of tert-butyl4-[acetyl(methoxy)amino]piperidine-1-carboxylate (110 mg, 0.40 mmol) inDCM (5 mL) was added TFA (2 mL) dropwise at 0° C. and the resultingmixture was stirred at rt for 3 h. The solvent was removed in vacuo andthe residue was dried by coevaporation from toluene (×3) to giveIntermediate 76, N-methoxy-N-(piperidin-4-yl)acetamide trifluoroacetatesalt (120 mg, 100%) as a gum which was used without furtherpurification.

The data for the title compound is in Table 2.

Route 13

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 89, 1-(1,3-oxazol-5-yl)methanaminehydrochloride

Intermediate 83, [(tert-butoxycarbonyl)amino]acetic acid (5.00 g, 28.5mmol), DIPEA (14.75 g, 104 mmol) and Intermediate 84,N-methoxymethanamine hydrochloride (5.60 g, 57.0 mmol) were dissolved inDCM (100 mL) and DMF (100 mL) and Intermediate 85, EDC hydrochloride(6.56 g, 34.0 mmol) was added. The reaction mixture was stirred undernitrogen at 0° C. for 1 h, then Intermediate 86, HOBt (4.63 g, 34.0mmol) and DMAP (100 mg) were added portionwise and the resulting mixturewas stirred for 16 h at room temperature. The reaction mixture waspartitioned between H₂O (250 mL) and DCM (100 mL), and the aqueous layerwas further extracted with EtOAc (2×100 mL). The combined organic layerswere dried (Na₂SO₄), filtered and the solvent was removed in vacuo. Theresidue was purified by column chromatography (Normal-Phase Silica, 0 to3% methanol in DCM) to give tert-butyl{2-[methoxy(methyl)amino]-2-oxoethyl}carbamate (4.50 g, 72%) as a solid.

LCMS (Method F): m/z 219 (M+H)⁺ (ES⁺), at 1.77 min, UV active.

tert-Butyl {2-[methoxy(methyl)amino]-2-oxoethyl}carbamate (4.50 g, 20.6mmol), was dissolved in THF (50.0 mL) and Intermediate 87, lithiumaluminium hydride (1.0 M in THF, 20.6 mL, 20.6 mmol) was added at −30°C. dropwise. The mixture was stirred for 20 min at −30° C., then excesssodium sulfate decahydrate was added portionwise. The mixture wasstirred for 30 min, then filtered through a Celite pad and the filtratewas concentrated in vacuo to give crude tert-butyl (2-oxoethyl)carbamate(390 mg, 85%) as a gum which was used without further purification.

¹H NMR: (400 MHz, DMSO-d₆) δ: 1.39-1.40 (m, 12H), 2.46-2.47 (m, 3H),2.89-2.95 (m, 1H), 3.05-3.12 (m, 1H), 4.11-4.19 (m, 1H).

tert-Butyl (2-oxoethyl)carbamate (3.00 g, 18.8 mmol), Intermediate 88,p-toluenesulfonylmethyl isocyanide (5.52 g, 28.2 mmol) and K₂CO₃ (7.78g, 56.4 mmol) were mixed in methanol (50 mL) and stirred at 0° C. over70 h. The reaction mixture was partitioned between H₂O (30 mL) and EtOAc(20 mL), and the aqueous layer was further extracted with EtOAc (2×20mL). The combined organic layers were dried (Na₂SO₄), filtered andconcentrated in vacuo. The residue was purified by column chromatography(Normal-Phase Silica, 0 to 3% MeOH in DCM) to give tert-butyl(1,3-oxazol-5-ylmethyl)carbamate (900 mg, 24%) as a gum.

LCMS (Method F): m/z 199 (M+H)⁺ (ES⁺), at 1.72 min, UV active.

tert-Butyl (1,3-oxazol-5-ylmethyl)carbamate (900 mg, 0.45 mmol) wasdissolved in 1,4-dioxane (10 mL) and 4 M HCl in 1,4-dioxane (10 mL) wasadded and the resulting mixture was stirred for 3 h at room temperature.The reaction mixture was concentrated and the residue was dried byco-evaporation from diethyl ether (5 mL) to give Intermediate 89,1-(1,3-oxazol-5-yl)methanamine hydrochloride salt (400 mg, 90%) as a gumwhich was used without further purification.

The data for the title compound are in Table 2

Route 14

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 98, methylpiperidin-4-yl(2,2,2-trifluoroethyl)carbamate hydrochloride

Intermediate 97, tert-butyl4-[(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (300 mg, 1.06mmol) was dissolved in acetonitrile (10 mL) and K₂CO₃ (450 mg, 3.19mmol) was added. The mixture was stirred at 70° C. for 2 h then cooledto 0° C. Intermediate 40, methyl carbonochloridate (0.12 mL, 1.59 mmol)was added dropwise and the resulting reaction mixture stirred at 25° C.for 8 h. The solvents were removed in vacuo and the residue waspartitioned between H₂O (120 mL) and EtOAc (100 mL). The aqueous layerwas further extracted with EtOAc (2×100 mL) and the combined organiclayers were dried (Na₂SO₄), filtered and concentrated in vacuo. Theresidue was purified by column chromatography (normal neutral activatedalumina, at 10% to 15% EtOAc in hexane) to give tert-butyl4-[(methoxycarbonyl)(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate(330 mg, 92%) as a gum.

LCMS (Method I): m/z 284 (M+H-56)⁺ (ES⁺), at 5.01 min, UV active.

tert-Butyl4-[(methoxycarbonyl)(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate(330 mg, 0.97 mmol) was dissolved in 1,4-dioxane (5 mL), 4.0 M HCl in1,4-dioxane (10 mL) was added dropwise and the resulting mixture wasstirred at 25° C. for 8 h. The solvents were removed in vacuo and theresidue was triturated with diethyl ether (3×3 mL) to give Intermediate98, methyl piperidin-4-yl(2,2,2-trifluoroethyl)carbamate hydrochloridesalt (210 mg, 90%) as a solid which was used without furtherpurification.

The data for the title compound are in Table 2.

Route 15

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 99, 2-methoxyethylpiperidin-4-yl(2,2,2-trifluoroethyl)carbamate hydrochloride

Intermediate 97, tert-butyl4-((2,2,2-trifluoroethyl)amino)piperidine-1-carboxylate (1.0 g, 3.50mmol) was dissolved in DMF (15 mL), Cs₂CO₃ (3.46 g, 10.6 mmol) and CuI(336 mg, 1.77 mmol) were added, and the reaction mixture was stirred at70° C. for 5 h then cooled to 25° C. Intermediate 22,1-bromo-2-methoxyethane (986 mg, 7.09 mmol) was added and the reactionmixture was stirred at 90° C. for 7 days. The solvents were removed invacuo and the residue was partitioned between H₂O (180 mL) and EtOAc(120 mL). The aqueous layer was further extracted with EtOAc (3×120 mL)and the combined organic layers were dried (Na₂SO₄), filtered andconcentrated in vacuo. The residue was purified by column chromatography(normal neutral activated alumina, at 8% EtOAc in hexane) to givetert-butyl4-{[(2-methoxyethoxy)carbonyl](2,2,2-trifluoroethyl)amino}piperidine-1-carboxylate(280 mg, 21%) as a gum.

LCMS (Method I): m/z 329 (M+H-56)⁺ (ES⁺), at 4.90 min, UV active.

tert-Butyl4-{[(2-methoxyethoxy)carbonyl](2,2,2-trifluoroethyl)amino}piperidine-1-carboxylate(240 mg, 0.71 mmol) was dissolved in 1,4-dioxane (5 mL), 4.0 M HCl in1,4-dioxane (10 mL) was added dropwise and the resulting reactionmixture was stirred at 25° C. for 8 h. The solvents were removed invacuo, and the residue was purified by triturating with diethyl ether(3×3 mL) to give Intermediate 99, 2-methoxyethylpiperidin-4-yl(2,2,2-trifluoroethyl)carbamate hydrochloride salt (170mg, 96%) as a solid.

The data for the title compound are in Table 2.

Route 16

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 109,N-cyclopropyl-N-(2,2,2-trifluoroethyl)piperidin-4-amine hydrochloride

Intermediate 11, tert-butyl 4-(cyclopropylamino)piperidine-1-carboxylate(200 mg, 0.83 mmol) was dissolved in THF (10 mL), Intermediate 107,N-methyl-2-pyrrolidinone (0.6 mL) and triethylamine (0.5 mL, 3.30 mmol)were added and the reaction mixture was stirred at 70° C. for 1 h thencooled to room temperature. Intermediate 108, 2,2,2-trifluoroethyltrifluoromethanesulfonate (385 mg, 1.66 mmol) was added and theresulting reaction mixture was stirred at 80° C. for 70 h. The solventswere removed in vacuo, and the residue was partitioned between H₂O (120mL) and EtOAc (100 mL) and the aqueous layer was further extracted withEtOAc (2×100 mL). The combined organic layers were dried (Na₂SO₄),filtered and concentrated in vacuo and the residue was purified bycolumn chromatography (normal neutral activated alumina, 0.5% MeOH inCH₂Cl₂) to give tert-butyl4-[cyclopropyl(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (220mg, 82%) as a gum.

LCMS (Method I): m/z 267 (M+H-56)⁺ (ES⁺), at 5.90 min, UV active.

tert-Butyl4-[cyclopropyl(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate (200mg, 0.62 mmol) was dissolved in 1,4-dioxane (5 mL), 4.0 M HCl in1,4-dioxane (5 mL) was added dropwise and the resulting reaction mixturewas stirred at 25° C. for 16 h. The solvents were removed in vacuo andthe residue was purified by triturating with diethyl ether (3×5 mL) togive Intermediate 109,N-cyclopropyl-N-(2,2,2-trifluoroethyl)piperidin-4-amine hydrochloridesalt (160 mg, 100%) as a solid.

The data for the title compound are in Table 2.

Route 17

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 111,2-[cyclopropyl(piperidin-4-yl)amino]ethanol trifluoroacetate

Intermediate 11, tert-butyl 4-(cyclopropylamino)piperidine-1-carboxylate(200 mg, 0.833 mmol) and cesium carbonate (0.812 g, 2.5 mmol) were addedto MeCN (10 mL) and stirred at 25° C. for 15 min. Intermediate 110,2-bromoethanol (134 mg, 1.08 mmol) was added and the reaction mixturewas stirred at 60° C. for 16 h. The reaction mixture was partitionedbetween water (30 mL) and 10% methanol in CH₂Cl₂ (30 mL) and the aqueouslayer was further extracted with 10% methanol in CH₂Cl₂ (2×30 mL). Theorganic layers were then combined, dried (Na₂SO₄), filtered andconcentrated to give tert-butyl4-[cyclopropyl(2-hydroxyethyl)amino]piperidine-1-carboxylate (180 mg,76%) as a gum which was used without further purification.

LCMS (Method F): m/z 285 (M+H)⁺ (ES⁺), at 1.46 min, UV active.

tert-Butyl 4-[cyclopropyl(2-hydroxyethyl)amino]piperidine-1-carboxylate(365 mg, 1.29 mmol) was dissolved in CH₂Cl₂ (10 mL), trifluoroaceticacid (1.1 mL, 12.9 mmol) was added and the resulting reaction mixturewas stirred at 25° C. for 16 h. The solvents were removed in vacuo andthe residue was purified by triturating with diethyl ether (3×10 mL) togive Intermediate 111, 2-[cyclopropyl(piperidin-4-yl)amino]ethanoltrifluoroacetate salt (380 mg, 100%) as a gum.

The data for the title compound are in Table 2.

Route 18

Typical Procedure for the Preparation of Amines, as Exemplified by thePreparation of Intermediate 112, tert-butyl4-(cyclobutylamino)piperidine-1-carboxylate

Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (500 mg, 2.51mmol) was dissolved in methanol (10 mL), Intermediate 9, cyclobutanamine(178 mg, 2.51 mmol), triethylamine (1.0 mL, 7.53 mmol) and ZnCl₂ (34 mg,0.25 mmol) were added at room temperature and the reaction mixture wasstirred at 60° C. for 4 h. NaBH₃CN (475 mg, 7.53 mmol) was addedportionwise, the resulting reaction mixture was stirred at 25° C. for 12h and then the solvents were removed in vacuo. The residue waspartitioned between H₂O (150 mL) and EtOAc (120 mL) and the aqueouslayer was further extracted with EtOAc (2×120 mL). The organic layerswere combined, dried (Na₂SO₄), filtered and concentrated in vacuo. Theresidue was purified by column chromatography (normal basic activatedalumina, at 10% to 30% EtOAc in hexane) to give Intermediate 112,tert-butyl 4-(cyclobutylamino)piperidine-1-carboxylate (560 mg, 88%) asa gum.

The data for the title compound are in Table 2.

Route 19

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 119,N-(2-methoxyethyl)-N-(2,2,2-trifluoroethyl)piperidin-4-aminetrifluoroacetate

Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (1.0 g, 5.02mmol), was dissolved in methanol (15 mL) and treated with Intermediate118, 2-methoxyethylamine (490 mg, 6.53 mmol), triethylamine (2.1 mL,15.1 mmol) and ZnCl₂ (68 mg, 0.50 mmol). The reaction mixture wasstirred at 65° C. for 7 h, then NaBH₃CN (949 mg, 15.1 mmol) was addedportionwise. The resulting reaction mixture was stirred at 25° C. for 17h. The solvents were removed in vacuo, and the residue was partitionedbetween H₂O (150 mL) and EtOAc (120 mL). The aqueous layer was extractedwith EtOAc (2×120 mL), and the organic layers were combined, dried(Na₂SO₄), and the solvent was removed in vacuo. The residue was purifiedby column chromatography (Normal basic activated alumina, 40% to 50%EtOAc in hexane) to give tert-butyl4-[(2-methoxyethyl)amino]piperidine-1-carboxylate (480 mg, 37%) as aliquid.

LCMS (Method I): m/z 203 (M+H-56)⁺ (ES⁺), at 3.60 min, UV active.

tert-Butyl 4-[(2-methoxyethyl)amino]piperidine-1-carboxylate (300 mg,1.16 mmol) was dissolved in THF (10 mL) and treated withN-methyl-2-pyrrolidinone (344 mg, 3.48 mmol) and triethylamine (0.7 mL,4.65 mmol). The reaction mixture was stirred at 70° C. for 1 h, thenIntermediate 108, 2,2,2-trifluoroethyl trifluoromethanesulfonate (297mg, 1.28 mmol) was added dropwise at 25° C. The resulting reactionmixture was stirred at 70° C. for 16 h. The solvents were removed invacuo and the reaction mixture was partitioned between H₂O (120 mL) andEtOAc (100 mL). The aqueous layer was further extracted with EtOAc(2×100 mL), and the combined organic layers were dried (Na₂SO₄). Thesolvent was removed in vacuo and residue was purified by columnchromatography (Normal neutral activated alumina, at 10% to % EtOAc inhexane) to give tert-butyl4-[(2-methoxyethyl)(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate(180 mg, 46%) as a gum.

LCMS (Method I): m/z 341 (M+H)⁺ (ES⁺), at 5.31 min, UV active.

tert-Butyl4-[(2-methoxyethyl)(2,2,2-trifluoroethyl)amino]piperidine-1-carboxylate(150 mg, 0.44 mmol) was dissolved in DCM (3 mL) and cooled to 0° C.Trifluoroacetic acid (0.8 mL) was added dropwise and the resultingreaction mixture was stirred at 25° C. for 8 h. The solvents wereremoved in vacuo and the residue was purified by triturating withdiethyl ether (3×2 mL) to give Intermediate 119,N-(2-methoxyethyl)-N-(2,2,2-trifluoroethyl)piperidin-4-aminetrifluoroacetate salt (105 mg, 67%) as a gum.

The data for the title compound are in Table 2

Route 20

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 120,N-ethyl-N-(oxetan-3-yl)piperidin-4-amine trifluoroacetate

Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (546 mg, 4.10mmol) was dissolved in methanol (20 mL). Intermediate 106,oxetan-3-amine (300 mg, 4.10 mmol), triethylamine (1.7 mL, 12.3 mmol)and ZnCl₂ (56 mg, 0.41 mmol) were added and then the reaction mixturewas stirred at 65° C. for 8 h. NaBH₃CN (776 mg, 1.23 mmol) was thenadded portionwise and the resulting reaction mixture was stirred at 25°C. for 17 h. The solvents were removed in vacuo, and the residue waspartitioned between H₂O (120 mL) and EtOAc (100 mL). The aqueous layerwas further extracted with EtOAc (2×100 mL) and the combined organiclayers were dried (Na₂SO₄). The solvent was removed in vacuo and theresidue was purified by triturating with pentane and decanting off thesolvents to give tert-butyl 4-(oxetan-3-ylamino)piperidine-1-carboxylate(680 mg, 97%) as a gum.

LCMS (Method I): m/z 257 (M+H)⁺ (ES⁺), at 2.92 min, UV active.

tert-Butyl 4-(oxetan-3-ylamino)piperidine-1-carboxylate (200 mg, 0.78mmol) was dissolved in methanol (10 mL) and Intermediate 20,acetaldehyde (103 mg, 2.34 mmol), triethylamine (0.3 mL, 2.34 mmol) andZnCl₂ (11 mg, 0.08 mmol) were added. The reaction mixture was stirred at50° C. for 7 h, then NaBH₃CN (148 mg, 2.34 mmol) was added portionwise.The resulting reaction mixture was stirred at 25° C. for 17 h. Thesolvents were removed in vacuo, and the residue was partitioned betweenH₂O (100 mL) and EtOAc (80 mL). The aqueous layer was further extractedwith EtOAc (2×80 mL), and the combined organic layers were dried(Na₂SO₄). The solvent was removed in vacuo and the residue was purifiedby column chromatography (Normal basic activated alumina, 0.5% to 3%MeOH in DCM) to give tert-butyl4-[ethyl(oxetan-3-yl)amino]piperidine-1-carboxylate (180 mg, 81%) as agum.

LCMS (Method I): m/z 285 (M+H)⁺ (ES⁺), at 3.84 min, UV active.

tert-Butyl 4-[ethyl(oxetan-3-yl)amino]piperidine-1-carboxylate (180 mg,0.63 mmol) was dissolved in DCM (8 mL) and cooled to 0° C.Trifluoroacetic acid (2 mL) was added dropwise and the resultingreaction mixture was stirred at 25° C. for 6 h. The solvents wereremoved in vacuo, and the residue was purified by triturating withdiethyl ether (3×1 mL) to give Intermediate 120,N-ethyl-N-(oxetan-3-yl)piperidin-4-amine trifluoroacetate salt (110 mg,95%) as a gum.

The data for the title compound are in Table 2

Route 21

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 128, N-ethyl-N-methoxypiperidin-4-aminetrifluoroacetate

To a stirred solution of Intermediate 75, O-methylhydroxylamine (0.5 g,6.0 mmol) in MeOH (25 mL) was added NaOAc (0.51 g, 6.2 mmol) and thereaction mixture was stirred at rt for five minutes. Intermediate 1,tert-butyl 4-oxopiperidine-1-carboxylate (1 g, 5.0 mmol), AcOH (0.5 g,8.8 mmol) and NaCNBH₃ (0.3 g, 5.0 mmol) were added and the reactionmixture was stirred at rt for 24 hours. The reaction mixture wasdistributed between EtOAc and water, and the EtOAc phase was separatedand washed with aq. NaHCO₃ and brine solution. The organic phase wasdried with Na₂SO₄ and concentrated in vacuo to give the crude product,which was purified by column chromatography (Normal-Phase Silica andEtOAc and hexanes as eluent solvents) to give tert-butyl4-(methoxyamino)piperidine-1-carboxylate (1 g, 90%) as a solid.

LCMS (Method F): m/z 231 (M+H)⁺ (ES⁺), at 2.07 min, UV active.

To a stirred solution of tert-butyl4-(methoxyamino)piperidine-1-carboxylate (300 mg, 1.30 mmol) in DMF (5ml) was added K₂CO₃ (540 mg, 3.91 mmol) and the mixture was stirred at80° C. for 1 hour. Intermediate 127, iodoethane (305 mg, 1.96 mmol) wasthen added and the mixture was stirred at 80° C. for 16 hours. Thereaction mixture was cooled to RT, diluted with cold water (10 ml) andthe compound was extracted with EtOAc (20 ml). The aqueous layer wasfurther extracted with EtOAc (2×20 mL) and the combined organic layerswere washed with brine solution then dried with Na₂SO₄. The solvent wasremoved in vacuo to give the crude product, which was purified by columnchromatography (Normal-Phase Silica and EtOAc and hexanes as eluentsolvents) to give tert-butyl4-[ethyl(methoxy)amino]piperidine-1-carboxylate (160 mg, 47%).

LCMS (Method F): m/z 259 (M+H)⁺ (ES⁺), at 2.02 min, UV active.

To a stirred solution of tert-butyl4-[ethyl(methoxy)amino]piperidine-1-carboxylate (160 mg, 0.62 mmol) inDCM (8 ml) was added TFA (3 mL) dropwise at 0° C. and the resultingmixture was stirred at room temperature for 3 h. The solvent was thenevaporated in vacuo and the residue was dried by co-evaporation fromtoluene (×3) to give Intermediate 128,N-ethyl-N-methoxypiperidin-4-amine trifluoroacetate salt (150 mg, 63%)as a gum.

The data for the title compound are in Table 2

Route 22

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 130,2-[phenyl(piperidin-4-yl)amino]ethanol hydrochloride

Intermediate 1, tert-butyl 4-oxopiperidine-1-carboxylate (3.00 g, 15.0mmol), Intermediate 77, aniline (1.40 g, 15.0 mmol), triethylamine (6.35mL, 45.0 mmol) and zinc chloride (0.75 mL, 0.75 mmol) were dissolved inmethanol (25.0 mL) under nitrogen and stirred at 50-60° C. for 16 h.NaCNBH₃ (2.84 g, 45.0 mmol) was added portionwise at 0-10° C. and theresulting mixture was stirred at 50-60° C. for 16 h. The reactionmixture was partitioned between H₂O (150 mL) and EtOAc (50 mL) and theaqueous layer was further extracted with EtOAc (2×50 mL). The combinedorganic layers were dried (Na₂SO₄) and the solvent was removed in vacuoto give the crude product, which was purified by column chromatography(Normal-Phase Silica, 0 to 3% methanol in DCM) to give tert-butyl4-(phenylamino)piperidine-1-carboxylate (1.30 g, 31%) as a solid.

LCMS (Method F): m/z 277 (M+H)⁺ (ES⁺), at 2.33 min, UV active.

tert-Butyl 4-(phenylamino)piperidine-1-carboxylate (350 mg, 1.26 mmol)and Intermediate 129, ethyl bromoacetate (274 mg, 1.64 mmol) weredissolved in DIPEA (3.0 mL) and stirred for 16 h at 90° C. The reactionmixture was partitioned between H₂O (30 mL) and EtOAc (20 mL) and theaqueous layer was further extracted with EtOAc (2×20 mL). The combinedorganic layers were dried (Na₂SO₄) and the solvent was removed in vacuoto give the crude product, which was purified by column chromatography(Normal-Phase Silica, 0 to 22% EtOAc in hexanes) to give tert-butyl4-[(2-ethoxy-2-oxoethyl)(phenyl)amino]piperidine-1-carboxylate (390 mg,85%) as a gum.

LCMS (Method F): m/z 363 (M+H)⁺ (ES⁺), at 2.72 min, UV active.

tert-Butyl4-[(2-ethoxy-2-oxoethyl)(phenyl)amino]piperidine-1-carboxylate (350 mg,0.96 mmol) was dissolved in THF (10.0 mL) and treated with lithiumborohydride solution in THF (3.0 M, 1.30 mL, 3.86 mmol) at 0° C. andstirred at room temperature for 48 h. The reaction mixture waspartitioned between cold aq. NH₄Cl solution (30 mL) and EtOAc (15 mL).The aqueous layer was further extracted with EtOAc (2×15 mL) and thecombined organic layers were dried (Na₂SO₄) and the solvent was removedin vacuo to give crude product, which was purified by columnchromatography (Normal-Phase Silica, 0 to 35% EtOAc in hexanes) to givetert-butyl 4-[(2-hydroxyethyl)(phenyl)amino]piperidine-1-carboxylate(270 mg, 87%) as a gum.

LCMS (Method F): m/z 321 (M+H)⁺ (ES⁺), at 1.94 min, UV active.

tert-Butyl 4-[(2-hydroxyethyl)(phenyl)amino]piperidine-1-carboxylate(265 mg, 0.82 mmol) was dissolved in 4 M HCl in 1,4-dioxane (5.0 mL) at0° C. and stirred at room temperature for 3 h. The reaction mixture wasconcentrated and then triturated with diethyl ether (3×10 mL) to giveIntermediate 130, 2-[phenyl(piperidin-4-yl)amino]ethanol hydrochloridesalt (200 mg, 94%) as a solid.

The data for the title compound are in Table 2.

Route 23

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 138,N-ethyl-N-[1-(piperidin-4-yl)propyl]acetamide trifluoroacetate

Intermediate 137, tert-butyl 4-propanoylpiperidine-1-carboxylate (450mg, 1.86 mmol), Intermediate 44, ethanamine (2.0 M solution in THF, 2.33mL, 4.66 mmol), Et₃N (0.780 mL, 5.60 mmol), ZnCl₂ (0.2 mL) and MeOH (10mL) were charged into a vial. The reaction mixture was heated at 60° C.for 4 h then the reaction mixture was cooled to 0° C. and NaCNBH₃ (351mg, 5.60 mmol) was added. The reaction mixture was allowed to warm toroom temperature and was stirred for 16 h. The reaction mixture wasconcentrated in vacuo and the residue was partitioned between H₂O (100mL) and EtOAc (100 mL). The aqueous layer was further extracted withEtOAc (2×50 mL) and the combined organic layers were dried (Na₂SO₄) andthe solvent was removed in vacuo to give the crude product, which waspurified by combi-flash column chromatography (Normal phase, Neutralsilica gel, 60-120 mesh, 0 to 1% MeOH in DCM) to give tert-butyl4-[1-(ethylamino)propyl]piperidine-1-carboxylate (440 mg, 87%) as a gum.

LCMS (Method F): m/z 271 (M+H)⁺ (ES⁺), at 5.34 min, UV active.

tert-Butyl 4-[1-(ethylamino)propyl]piperidine-1-carboxylate (435 mg,1.61 mmol) was dissolved in DCM (10 mL) and triethylamine (0.67 mL, 4.83mmol) was added dropwise at 0-5° C. The reaction mixture was stirred at0-5° C. for 10 min then acetyl chloride (0.17 mL, 2.41 mmol) was addeddropwise at 0-5° C. The resulting reaction mixture was stirred at 25° C.for 8 h, then the solvents were removed in vacuo. The residue waspartitioned between H₂O (50 mL) and DCM (50 mL) and the aqueous layerwas further extracted with DCM (2×30 mL). The combined organic layerswere dried (Na₂SO₄) and the solvents were removed in vacuo. The residuewas purified by column chromatography (Normal basic activated alumina,at 0.5% to 1.0% MeOH in DCM) to give tert-butyl4-{1-[acetyl(ethyl)amino]propyl}piperidine-1-carboxylate (415 mg, 63%)as a gum.

LCMS (Method I): m/z 313 (M+H)⁺ (ES⁺), at 4.53 min, UV active.

tert-Butyl 4-{1-[acetyl(ethyl)amino]propyl}piperidine-1-carboxylate (415mg, 1.33 mmol) was dissolved in DCM (5.0 mL) at 0° C. and TFA (2.5 mL)was added to the solution at 0° C. The reaction mixture was allowed towarm at room temperature and was stirred for 6 h. The reaction mixturewas then concentrated and dried by co-evaporation from diethyl ether(3×5 mL) to give Intermediate 138,N-ethyl-N-[1-(piperidin-4-yl)propyl]acetamide trifluoroacetate salt (250mg, 80%) as a gum.

The data for the title compound are in Table 2.

Route 24

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 140,N-ethyl-N-[2-(piperidin-4-yl)propan-2-yl]acetamide hydrochloride

Intermediate 139, tert-butyl4-(2-aminopropan-2-yl)piperidine-1-carboxylate (300 mg, 1.24 mmol),Intermediate 20, acetaldehyde (163 mg, 3.71 mmol), triethylamine (0.52mL, 3.71 mmol) and zinc chloride (0.06 mL, 0.06 mmol) were dissolved inmethanol (10 mL) under nitrogen and stirred for 16 h at 50-60° C. After16 h, NaCNBH₃ (233 mg, 3.74 mmol) was added portionwise at 0-10° C. andthe resulting mixture was stired for 6 h at 50-60° C. The reactionmixture was partitioned between H₂O (40 mL) and EtOAc (25 mL) and theaqueous layer was further extracted with EtOAc (2×25 mL). The combinedorganic layers were dried (Na₂SO₄) and the solvent was removed in vacuoto give the crude product, which was purified by column chromatography(Normal-Phase Silica, 0 to 4% MeOH in DCM) to give tert-butyl4-[2-(ethylamino)propan-2-yl]piperidine-1-carboxylate (180 mg, 54%) as agum.

LCMS (Method F): m/z 271 (M+H)⁺ (ES⁺), at 1.71 min, UV active.

tert-butyl 4-[2-(ethylamino)propan-2-yl]piperidine-1-carboxylate (180mg, 0.66 mmol) and triethylamine (0.27 mL, 19.9 mmol) were dissolved indry DCM (5.0 mL) under nitrogen. Acetyl chloride (78.0 mg, 0.99 mmol)was added at 0° C. and the resulting mixture was stirred for 30 min atroom temperature. The reaction mixture was then partitioned between sat.aq. NaHCO₃ solution (20 mL) and EtOAc (15 mL) and the aqueous layer wasfurther extracted with EtOAc (2×25 mL). The combined organic layers weredried (Na₂SO₄) and the solvent was removed in vacuo to give the crudeproduct, which was purified by column chromatography (Normal-PhaseSilica, 0 to 65% EtOAc in hexanes) to give tert-butyl4-{2-[acetyl(ethyl)amino]propan-2-yl}piperidine-1-carboxylate (160 mg,77%) as a gum.

LCMS (Method I): m/z 257 (M+H-56)⁺ (ES⁺), at 4.75 min, UV active.

tert-butyl 4-{2-[acetyl(ethyl)amino]propan-2-yl}piperidine-1-carboxylate(160 mg, 0.51 mmol) and 4 M HCl in 1,4-dioxane (5 mL) were dissolved in1,4-dioxane (5 mL) under nitrogen and stirred together for 3 h at roomtemperature. The reaction mixture was poured into toluene and thentriturated with diethyl ether (2×5 mL) and concentrated in vacuo to giveIntermediate 140, N-ethyl-N-[2-(piperidin-4-yl)propan-2-yl]acetamidehydrochloride salt (110 mg, 95%) as a solid.

The data for the title compound are in Table 2.

Route 25

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 144,4-[1-(1H-pyrazol-1-yl)ethyl]piperidine trifluoroacetate

Intermediate 141, tert-butyl 4-(1-hydroxyethyl)piperidine-1-carboxylate(2.0 g, 8.73 mmol) and Et₃N (3.64 mL, 26.3 mmol) were dissolved indichloromethane (20.0 mL) and cooled to 0° C. Intermediate 142,methanesulfonyl chloride (0.82 mL, 10.4 mmol) was added dropwise and thereaction mixture was allowed to stir at 0° C. for 2 h. The reactionmixture was diluted with water (100 mL) and extracted with DCM (2×30mL). The combined organic layers were dried (Na₂SO₄) and concentrated invacuo to give crude tert-butyl4-{1-[(methylsulfonyl)oxy]ethyl}piperidine-1-carboxylate (2.0 g, 75%) asan oil. The crude product was used in the next step without furtherpurification.

LCMS (Method I): m/z 252 (M+H-56)⁺ (ES⁺), at 4.51 min, UV active.

Intermediate 143, 1H-pyrazole (887 mg, 13.03 mmol) was dissolved in DMF(15.0 mL) and cooled to 0° C. 60% sodium hydride suspension in mineraloil (281 mg, 7.0 mmol) was added and the mixture was allowed to stir at0° C. for 1 h. After the completion of 1 h, tert-butyl4-{1-[(methylsulfonyl)oxy]ethyl}piperidine-1-carboxylate (2.0 g, 6.51mmol) in DMF (1.0 mL) was added dropwise at 0° C. and the reactionmixture was stirred at room temperature for 16 h. The mixture wasdiluted with water (100 mL) and extracted with DCM (2×30 mL). Thecombined organic layers were dried (Na₂SO₄) and concentrated in vacuo togive the crude product, which was purified by column chromatography(Normal phase, Neutral silica gel, 60-120 mesh, 0 to 30% EtOAc inhexane) to give tert-butyl4-[1-(1H-pyrazol-1-yl)ethyl]piperidine-1-carboxylate (430 mg, 24%) as agum.

LCMS (Method I): m/z 280 (M+H)⁺ (ES⁺), at 4.51 min, UV active.

tert-Butyl 4-[1-(1H-pyrazol-1-yl)ethyl]piperidine-1-carboxylate (430 mg,1.54 mmol) was dissolved in dichloromethane (8.0 mL) and cooled to 0° C.TFA (4.0 mL) was added and the reaction mixture was stirred at roomtemperature for 2 h. The reaction mixture was then concentrated in vacuoto give crude Intermediate 144, 4-[1-(1H-pyrazol-1-yl)ethyl]piperidinetrifluoroacetate salt (450 mg, 100%) as a gum, which was used withoutfurther purification. The data for the title compound are in Table 2.

Route 26

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 147, 4-(1-phenylethoxy)piperidinehydrochloride

Intermediate 145, tert-butyl 4-hydroxypiperidine-1-carboxylate (543 mg,2.69 mmol) was dissolved in DMF (10 mL), 60% sodium hydride suspensionin mineral oil (183 mg, 4.58 mmol) was added portionwise under nitrogenat 0° C. and the mixture was stirred at room temperature for 1 h. After1 h, Intermediate 146, (1-bromoethyl)benzene (500 mg, 2.69 mmol) wasadded dropwise and the resulting mixture was stirred for 16 h at 90° C.The reaction mixture was partitioned between H₂O (50 mL) and EtOAc (25mL) and the aqueous layer was further extracted with EtOAc (2×25 mL).The organic layers were combined, dried (Na₂SO₄) and the solvent wasremoved in vacuo to give the crude product, which was purified by columnchromatography (Normal-Phase Silica, 0 to 15% EtOAc in hexanes) to givetert-butyl 4-(1-phenylethoxy)piperidine-1-carboxylate (161 mg, 20%) as agum.

LCMS (Method F): m/z 306 (M+H)⁺ (ES⁺), at 2.79 min, UV active.

tert-Butyl 4-(1-phenylethoxy)piperidine-1-carboxylate (160 mg, 5.27mmol) was dissolved in 4 M HCl in 1,4-dioxane (5 mL) at 0° C. andstirred at room temperature for 3 h. The reaction mixture wasconcentrated and the residue was triturated with diethyl ether (3×10 mL)to give Intermediate 147, 4-(1-phenylethoxy)piperidine hydrochloridesalt (100 mg, 89%) as a solid.

The data for the title compound are in Table 2.

Route 27

Typical Procedure for the Preparation of Piperidines, as Exemplified bythe Preparation of Intermediate 150, 4-(benzylsulfanyl)piperidinehydrochloride

Intermediate 149, phenylmethanethiol (9.6 mL, 81.8 mmol) was dissolvedin DMF (80 mL), 60% sodium hydride suspension in mineral oil (3.27 g,81.8 mmol) was added portionwise under nitrogen at 0° C. and theresulting mixture was stirred at room temperature for 30 min. After 30min, Intermediate 148, tert-butyl 4-bromopiperidine-1-carboxylate (5.4g, 20.4 mmol) was added dropwise and the resulting mixture was stirredfor 16 h at room temperature. The reaction mixture was partitionedbetween H₂O (150 mL) and EtOAc (50 mL) and the aqueous layer was furtherextracted with EtOAc (2×50 mL). The combined organic layers were dried(Na₂SO₄) and the solvent was removed in vacuo to give the crude product,which was purified by column chromatography (Normal-Phase Silica, 0 to12% EtOAc in hexanes) to give tert-butyl4-(benzylsulfanyl)piperidine-1-carboxylate (1.59 g, 25%) as a gum.

LCMS (Method F): m/z 252 (M+H-56)⁺ (ES⁺), at 2.73 min, UV active.

tert-Butyl 4-(benzylsulfanyl)piperidine-1-carboxylate (1.00 g, 3.25mmol) was dissolved in 4 M HCl in 1,4-dioxane (10 mL) at 0° C. andstirred at room temperature for 3 h. The reaction mixture wasconcentrated and the residue was triturated with diethyl ether (3×10 mL)to give Intermediate 150, 4-(benzylsulfanyl)piperidine hydrochloridesalt (750 mg, 95%) as a gum. The data for the title compound are inTable 2.

Route 28

Procedure for the Preparation of Intermediate 152, methyl6-oxo-2-azaspiro[3.4]octane-2-carboxylate

Intermediate 151, tert-butyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate(120 mg, 0.533 mmol) was dissolved in DCM (2.0 mL) at 0° C. and TFA (1.0mL) was added. The reaction mixture was allowed to warm to roomtemperature and was stirred for 2 h, then concentrated in vacuo. Theresidue was dried by co-evaporation from diethyl ether (3×10 mL) to give2-azaspiro[3.4]octan-6-one trifluoroacetate salt (120 mg, 100%) as agum.

LCMS (Method I): m/z 125 (M+H)⁺ (ES⁺), at 0.60 min, UV active.

2-Azaspiro[3.4]octan-6-one trifluoroacetate salt (60 mg, 0.251 mmol) wasdissolved in DCM (5 mL) and triethylamine (0.2 mL, 1.25 mmol) was addedat 0° C. Intermediate 40, methyl carbonochloridate (94.4 mg, 0.37 mmol)was added at 0° C. and the reaction mixture was allowed to warm to roomtemperature and was stirred for 2 h. The mixture was concentrated invacuo and the residue was partitioned between H₂O (25 mL) and EtOAc (25mL). The aqueous layer was further extracted with EtOAc (2×10 mL) andthe combined organic layers were dried (Na₂SO₄) and the solvent wasremoved in vacuo to give Intermediate 152, methyl6-oxo-2-azaspiro[3.4]octane-2-carboxylate (30 mg, 34%) as an oil.

The data for the title compound are in Table 2.

General Synthetic Procedures for the Examples

Route a

Typical Procedure for the Preparation of Piperidines Via ReductiveAmination, as Exemplified by the Preparation of Example 2-2, Ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 2, N-ethyl-N-(piperidin-4-yl)acetamide hydrochloride (150mg, 0.9 mmol), Intermediate 4, ethyl2-oxo-6-azaspiro[3.4]octane-6-carboxylate (170 mg, 0.9 mmol), Et₃N (0.24mL, 1.9 mmol) and ZnCl₂ (6 mg) were dissolved in MeOH (10 mL) and thereaction mixture was stirred at 60° C. for 8 h. The mixture was thencooled to 0° C. and NaBH₃CN (72 mg, 1.2 mmol) was added portionwise,after which the mixture was stirred at 25° C. for 17 h. The solvent wasremoved in vacuo, and the residue was partitioned between H₂O (100 mL)and EtOAc (80 mL). The aqueous layer was further extracted with EtOAc(2×80 mL) and the combined organic phases were dried (Na₂SO₄) and thesolvent was removed in vacuo. The residue was purified by prep. HPLC[reverse phase (X-BRIDGE C-18, 150×19 mm, 5 μm, 15 mL per min, gradient25% to 100% (over 20 min), 100% (over 3 min), then 30% (over 2 min),0.1% NH₃ in MeCN/water] to give Example 2-2 Isomer 1, ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(30 mg, 9%) as a gum and Example 2-2 Isomer 2, ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(25 mg, 7%) as a gum.

The data for Example 2-2 Isomer 2 are in Table 3.

Route b

Typical Procedure for the Preparation of Piperidines Via SodiumTriacetoxyborohydride Reductive Amination as Exemplified by thePreparation of Example 2-12, ethyl2-(4-{[acetyl(cyclopropyl)amino]methyl}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 4, ethyl 2-oxo-6-azaspiro[3.4]octane-6-carboxylate (99 mg,0.5 mmol) and Intermediate 35,N-cyclopropyl-N-(piperidin-4-ylmethyl)acetamide hydrochloride (116 mg,0.5 mmol) were dissolved in DCM (10 mL) at rt and Et₃N (0.35 mL, 2.5mmol) was added. The mixture was stirred for 30 min before addition ofAcOH (0.29 mL, 5.0 mmol). The mixture was stirred for 3 h, then STAB(265 mg, 1.3 mmol) was added and the mixture was stirred at rtovernight. The reaction mixture was quenched with the addition of sat.aq. NaHCO₃ (20 mL), and solid Na₂CO₃ was added to ensure the aqueouslayer was basic. The resulting mixture was extracted with DCM (4×20 mL)and the organic layers were combined, dried (MgSO₄), filtered and thesolvents were removed in vacuo. The residue was purified by columnchromatography (normal phase, [Biotage SNAP cartridge KP-sil 10 g, 40-63μm, 60 Å, 20 mL per min, gradient 0% to 10% MeOH in DCM]) to give aninseparable mixture of diastereoisomers. This mixture was purified bypreparative reversed phase HPLC (Phenomenex Gemini-NX 5 μm C18 110 AAxia column, 100×30 mm, eluting with 30 to 60% MeCN/Solvent B over 14.4min at 30 mL/min [where solvent B is 0.2% of (28% NH₃/H₂O) in H₂O] andcollecting fractions by monitoring at 205 nm) to give Example 2-12Isomer 1, ethyl2-(4-{[acetyl(cyclopropyl)amino]methyl}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate(42 mg, 22%) as a solid and Example 2-12 Isomer 2 ethyl2-(4-{[acetyl(cyclopropyl)amino]methyl}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate(36 mg, 19%) as a solid.

The data for Example 2-12 Isomer 2 are in Table 3.

Route c

Typical Procedure for the Preparation of Piperidines Via Use of aProtected Ketone as Exemplified by the Preparation of Example 2-23,ethyl2-(4-{acetyl[(3-methyloxetan-3-yl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 57, 1,4-dioxa-8-azaspiro[4.5]decane (1.0 g, 6.99 mmol) wasdissolved in methanol (20 mL) and Intermediate 4, ethyl2-oxo-6-azaspiro[3.4]octane-6-carboxylate (1.38 g, 6.99 mmol),triethylamine (2.9 mL, 20.9 mmol) and ZnCl₂ (95 mg, 0.70 mmol) wereadded and then the reaction mixture was stirred at 65° C. for 8 h.NaBH₃CN (1.32 g, 20.9 mmol) was added portionwise and the resultingreaction mixture was stirred at 25° C. for 17 h. The solvents wereremoved in vacuo, and the residue was partitioned between H₂O (120 mL)and EtOAc (100 mL). The aqueous layer was further extracted with EtOAc(2×100 mL), the organic layers were combined, dried (Na₂SO₄) and thesolvent was removed in vacuo. The residue was purified by trituratingwith pentane and decanting off the solvents to give ethyl2-(1,4-dioxa-8-azaspiro[4.5]dec-8-yl)-6-azaspiro[3.4]octane-6-carboxylate(1.80 g, 79%) as a gum.

LCMS (Method I): m/z 325 (M+H)⁺ (ES⁺), at 3.54 and 3.69 min, UV active.

Ethyl2-(1,4-dioxa-8-azaspiro[4.5]dec-8-yl)-6-azaspiro[3.4]octane-6-carboxylate(1.80 μg, 5.55 mmol) was dissolved in ethanol (20 mL) and 4.0 M HCl in1,4-dioxane (30 mL) was added. The resulting reaction mixture wasstirred at 70° C. for 18 h and then basified with sat. sodiumbicarbonate solution. The solvents were removed in vacuo and the residuewas partitioned between H₂O (100 mL) and EtOAc (80 mL). The aqueouslayer was further extracted with EtOAc (2×80 mL) and the combinedorganic layers were dried (Na₂SO₄). The solvent was removed in vacuo andthe residue was triturated with pentane to give ethyl2-(4-oxopiperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate (1.20 g,77%) as a gum.

LCMS (Method I): m/z 281 (M+H)⁺ (ES⁺), at 3.30 and 3.41 min, UV active.

Intermediate 58, 1-(3-methyloxetan-3-yl)methanamine (72 mg, 0.72 mmol),ethyl 2-(4-oxopiperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate (200mg, 0.71 mmol), triethylamine (0.4 mL, 2.85 mmol) and ZnCl₂ (9 mg, 0.07mmol) were dissolved in MeOH (10 mL) and the reaction mixture wasstirred at 65° C. for 8 h. The mixture was cooled to 0° C. and NaBH₃CN(134 mg, 2.14 mmol) was added portionwise. The resulting reactionmixture was stirred at 25° C. for 17 h. The solvents were removed invacuo and the residue was partitioned between H₂O (80 mL) and EtOAc (60mL). The aqueous layer was further extracted with EtOAc (2×60 mL) andthe combined organic layers were dried (Na₂SO₄) and the solvent wasremoved in vacuo. The residue was purified by triturating with pentane(3×1 mL) to give ethyl2-(4-{[(3-methyloxetan-3-yl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate(210 mg, 81%) as a gum.

LCMS (Method I): m/z 366 (M+H)⁺ (ES⁺), at 3.63 and 3.81 min, UV active.

Ethyl2-(4-{[(3-methyloxetan-3-yl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate(200 mg, 0.55 mmol) was dissolved in DCM (10 mL), triethylamine (0.2 mL,1.64 mmol) was added and the reaction mixture was stirred at 0° C. for20 min. Acetyl chloride (0.06 mL, 0.82 mmol) was added portionwise andthe resulting reaction mixture was stirred at 25° C. for 2 h. Thesolvents were removed in vacuo, the residue was partitioned between H₂O(80 mL) and EtOAc (60 mL) and the aqueous layer was further extractedwith EtOAc (2×60 mL). The combined organic layers were dried (Na₂SO₄),the solvent was removed in vacuo and the residue was purified bypreparative HPLC [reverse phase (X-BRIDGE C18, 250×19 mm, 5 μm, 15 mLper min, gradient 5% to 30% (over 36 min), 30% (over 9 min), 100% (over5 min), then 5% (over 5 min), mobile phase (A) 5 mM ammoniumbicarbonate+0.1% ammonia in water and (B) 100% acetonitrile] to giveExample 2-23 Isomer 1, ethyl2-(4-{acetyl[(3-methyloxetan-3-yl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate(47 mg, 21%) as a liquid and Example 2-23 Isomer 2, ethyl2-(4-{acetyl[(3-methyloxetan-3-yl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate(45 mg, 20%) as a liquid.

The data for Example 2-23 Isomer 2 are in Table 3.

Route d

Typical Procedure for the Preparation of Piperidines Via NucleophilicDisplacement on a 4-nitrophenyl carbamate as Exemplified by thePreparation of Example 2-38, (1,1-²H₂)-ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 114, N-ethyl-N-(piperidin-4-yl)acetamide (1.70 g, 10 mmol)and Intermediate 3, tert-butyl 2-oxo-6-azaspiro[3.4]octane-6-carboxylate(2.25 g, 10 mmol) were mixed in DMF (40 mL) under an atmosphere ofnitrogen. AcOH (0.86 mL, 15 mmol) and STAB (4.24 g, 20 mmol) were addedand the resulting mixture was stirred at rt for 6 d. The mixture wasconcentrated to remove DMF and the residue was treated with toluene andconcentrated to remove AcOH. The residue was dissolved in MeOH andconcentrated onto flash silica (15 mL). The resulting powder waspurified by column chromatography (normal phase, [Biotage SNAP cartridgeKP-sil 100 g, 40-63 μm, 60 Å, 40 mL per min, gradient 0% to 10% SolventA in DCM over 15 CV, where Solvent A is 10% of {7 M NH₃ in MeOH} inMeOH]) to give tert-butyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateas a gum (2.92 g, 77%).

LCMS (Method D): m/z 380 (M+H)⁺ (ES⁺), at 2.11 min, UV active.

tert-Butyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(2.80 g, 7.38 mmol) was dissolved in a mixture of DCM (50 mL) and TFA(50 mL) under an atmosphere of nitrogen and stirred ar rt for 3.5 h. Themixture was diluted with toluene and concentrated. The oily residue wasdiluted with toluene and concentrated to affordN-[1-(6-azaspiro[3.4]oct-2-yl)piperidin-4-yl]-N-ethylacetamidetrifluoroacetate salt as a gum (5.73 g, assumed 100%).

LCMS (Method D): m/z 280 (M+H)⁺ (ES⁺), at 1.67 and 1.79 min, weakly UVactive.

N-[1-(6-Azaspiro[3.4]oct-2-yl)piperidin-4-yl]-N-ethylacetamidetrifluoroacetate salt (5.73 g, assumed 7.38 mmol) was dissolved in DCM(140 mL) under an atomosphere of nitrogen. Et₃N (5.1 ml, 36.6 mmol) andIntermediate 115, 4-nitrophenyl carbonochloridate (1.78 g, 8.83 mmol)were added and the resulting mixture was stirred at rt overnight. MoreEt₃N (2 ml, 14.3 mmol) and Intermediate 115, 4-nitrophenylcarbonochloridate (0.74 g, 3.67 mmol) were added and the mixture wasstirred at rt for a further 3 d. The reaction mixture was concentratedonto flash silica (15 mL) and the resulting powder was purified bycolumn chromatography (normal phase, [Biotage SNAP cartridge KP-sil 100g, 40-63 μm, 60 Å, 40 mL per min, gradient 0% to 5% Solvent A in DCMover 10 CV, where Solvent A is 10% of {7 M NH₃ in MeOH} in MeOH]) togive an oil which was further purified by column chromatography (normalphase, [Biotage SNAP cartridge KP-sil 100 g, 40-63 μm, 60 Å, 40 mL permin, isochratic 5% EtOAc in DCM over 5 CV then isochratic 5% Solvent Ain DCM over 5 CV, where Solvent A is 10% of {7 M NH₃ in MeOH} in MeOH])to give an oil (6.85 g). The oil was dissolved in DCM, washed with H₂O(×2), dried by passing through a phase separator cartridge andconcentrated to give 4-nitrophenyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateas a foam (2.41 g, 73%).

LCMS (Method C): m/z 445 (M+H)⁺ (ES⁺), at 1.32 min, UV active.

Intermediate 116, (1,1-²H₂)-ethanol (0.42 mL, 7.19 mmol) was dissolvedin THF (18 mL) under an atmosphere of nitrogen and treated with 60%sodium hydride suspension in mineral oil (0.29 g, 7.25 mmol). Themixture was stirred at rt for 1 h, then 4-nitrophenyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(0.800 g, 1.80 mmol) was added and the resulting mixture was stirred atrt for 4 d. The reaction mixture was concentrated to remove THF then theresidue was diluted with H₂O and extracted with EtOAc (×2). The combinedorganic phases were passed through a phase separator cartidge andconcentrated onto flash silica (10 mL). The resulting powder waspurified by column chromatography (normal phase, [Biotage SNAP cartridgeKP-sil 50 g, 40-63 μm, 60 Å, 40 mL per min, isochratic 20% EtOAc in DCMover 5 CV, gradient 20% EtOAc in DCM to 10% Solvent A in DCM over 1 CV,isochratic 10% Solvent A in DCM over 10 CV, where Solvent A is 10% of {7M NH₃ in MeOH} in MeOH]) to give an inseparable mixture ofdiastereoisomers (0.359 g, 56%). This mixture was purified bypreparative reversed phase HPLC (Phenomenex Gemini-NX 5 μm C18 110 AAxia column, 100×30 mm, eluting with 20 to 50% MeCN/Solvent B over 14.4min at 30 mL/min [where solvent B is 0.2% of (28% NH₃/H₂O) in H₂O] andcollecting fractions by monitoring at 205 nm) to give Example 2-38Isomer 1, (1,1-²H₂)-ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(140 mg, 22%) as a gum and Example 2-38 Isomer 2, (1,1-²H₂)-ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(129 mg, 20%) as a gum.

The data for Example 2-38 Isomer 2 are in Table 3.

Route e

Typical Procedure for the Preparation of Piperidines where a Mixture ofMore than Two Isomers is Separated by Using Reversed PhaseChromatography Followed by Chiral Chromatography as Exemplified by thePreparation of Example 2-63, ethyl2-(4-{1-[acetyl(ethyl)amino]propyl}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 138, N-ethyl-N-[1-(piperidin-4-yl)propyl]acetamidetrifluoroacetate (250 mg, 1.18 mmol), Intermediate 4, ethyl2-oxo-6-azaspiro[3.4]octane-6-carboxylate (232 mg, 1.18 mmol), Et₃N(0.821 mL, 5.89 mmol), ZnCl₂ (0.3 mL) and MeOH (5 mL) were charged intoa vial. The resulting mixture was heated at 60° C. for 4 h then cooledto 0° C. NaCNBH₃ (222 mg, 3.53 mmol) was added at 0° C. and the mixturewas allowed to warm to room temperature and stirred for 16 h. Themixture was concentrated in vacuo and the residue was partitionedbetween H₂O (50 mL) and EtOAc (50 mL). The aqueous layer was furtherextracted with EtOAc (2×50 mL) and the combined organic layers weredried (Na₂SO₄) and the solvent was removed in vacuo to give the crudeproduct, which was purified by preparative HPLC [reverse phase HPLC(X-BRIDGE, 250×19 mm, 5 μm, 15 mL per min, gradient 48% (over 60 min),100% (over 2 min) then 48% (over 3 min), (A) 10 mM ammonium bicarbonatein water+0.1% NH₃ in water, (B) 50:50 (MeCN:MeOH)] to give twoisomers—isomer 1 and isomer 2.

The two isomers were taken in turn and further purified by chiralpreparative HPLC [CHIRALCEL OX-H 250×4.6 mm, 5 μm {0.3% DEA in IPA:MeOH(50:50)} to give Example 2-63 Isomer 1a, ethyl2-{4-[(2-hydroxyethyl)(phenyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(13 mg, 3%) as a gum, Example 2-63 Isomer 1b, ethyl2-{4-[(2-hydroxyethyl)(phenyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(10 mg, 2%) as a gum, Example 2-63 Isomer 2a, ethyl2-{4-[(2-hydroxyethyl)(phenyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(11 mg, 2%) as a gum and Example 2-63 Isomer 2b, ethyl2-{4-[(2-hydroxyethyl)(phenyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(10 mg, 2%) as a gum.

The data for Example 2-63 Isomer 2b are in Table 3.

Route f

Typical Procedure for the Preparation of Piperidines where a Mixture ofMore than Two Isomers is Separated by Using Reversed PhaseChromatography Followed by Chiral Chromatography as Exemplified by thePreparation of Example 2-65, ethyl2-{4-[1-(1H-pyrazol-1-yl)ethyl]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate

Intermediate 144, 4-[1-(1H-pyrazol-1-yl)ethyl]piperidinetrifluoroacetate salt (430 mg, 2.40 mmol) and Et₃N (1.6 mL, 12.0 mmol)were dissolved in methanol (10 mL). Intermediate 4, ethyl2-oxo-6-azaspiro[3.4]octane-6-carboxylate (473 mg, 2.40 mmol) and ZnCl₂(0.12 mL, 0.12 mmol) were added and the reaction mixture was stirred at70° C. for 5 h. The reaction mixture was cooled to 0° C. and NaCNBH₃(452 mg, 7.21 mmol) was added and the reaction mixture was stirred atroom temperature for 16 h. The reaction mixture was concentrated invacuo, diluted with water (30 mL) and extracted with ethyl acetate (2×10mL). The combined organic layers were washed with brine, dried overNa₂SO₄ and concentrated in vacuo. The crude product was purified bypreparative HPLC [X-BRIDGE C18 (250×19 mm) 5 μm, 12 mL per min, gradient30% to 100% (over 120 min) then 100% (5 min) [5 mM ammonium bicarbonatein water/MeCN:MeOH (50:50)] to give two isomers—isomer 1 and isomer 2.

Isomer 1 was further purified by Chiral SFC [Chiral CEL OX-H (250×4.6mm) 5 μm, co-solvent: 15% of 0.3% DEA in IPA:MeOH (50:50) to giveExample 2-65 Isomer 1a, ethyl2-{4-[1-(1H-pyrazol-1-yl)ethyl]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(17 mg, 28%) and Example 2-65 Isomer 1b, ethyl2-{4-[1-(1H-pyrazol-1-yl)ethyl]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(25 mg, 42%).

The data for Example 2-65 Isomer 1a are in Table 3.

Isomer 2 was further purified by Chiral SFC [Chiral PAK ADH (250×4.6 mm)5 μm, co-solvent: 35% of 0.3% DEA in MeOH to give Example 2-65 Isomer2a, ethyl2-{4-[1-(1H-pyrazol-1-yl)ethyl]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(19 mg, 32%) and Example 2-65 Isomer 2b, ethyl2-{4-[1-(1H-pyrazol-1-yl)ethyl]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate(21 mg, 35%).

The data for Example 2-65 Isomer 2a are in Table 3.

Route p

Typical Procedure for the Preparation of Piperidines Containing the2-azaspiro[3.4]octane Ring System where a Mixture of Two Enantiomers isSeparated by Using by Using Reversed Phase Chromatography Followed byChiral Chromatography as Exemplified by the Preparation of Example 3-2,methyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylate

Intermediate 2, N-ethyl-N-(piperidin-4-yl)acetamide hydrochloride (150mg, 0.818 mmol), Intermediate 152, methyl6-oxo-2-azaspiro[3.4]octane-2-carboxylate (139 mg, 0.818 mmol),triethylamine (0.342 mL, 2.45 mmol) and ZnCl₂ (1.0 M solution in diethylether, 0.2 mL, 0.2 mmol) were dissolved in MeOH (100 mL) and the mixturewas stirred at 60° C. for 8 h. The mixture was then cooled down to 0-5°C. and NaBH₃CN (154 mg, 2.45 mmol) was added portionwise. The resultingreaction mixture was stirred at 25° C. for 17 h, then the solvents wereremoved in vacuo. The residue was partitioned between H₂O (100 mL) andEtOAc (100 mL) and the aqueous layer was further extracted with EtOAc(2×50 mL). The combined organic layers were dried (Na₂SO₄) and thesolvent was removed in vacuo. The residue was purified by prep. HPLC[reverse phase (PHENYL HEXYL, 250×19 mm, 5 μm, 14 mL per min, gradient35% (over 9 min), 100% (over 2 min), then 35% (over 2 min), A: 0.1%ammonia in water, B: 100% MeCN] to give methyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylate(65 mg, 22%) as a gum. This was further purified by chiral prep. HPLC(CHIRALPAK AD-H 250×4.6 mm, 5 μm, co-solvent 0.3% diethylamine in MeOH)to give Example 3-2 Isomer 1, methyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylate(27 mg, 9%) as a gum and Example 3-2 Isomer 2, methyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylate(31 mg, 11%) as a gum. The data for Example 3-2 Isomer 2 are in Table 3.

TABLE 2 Characterising data and commercial sources for startingmaterials and intermediates Table 2 Intermediate Route Name Data 1tert-butyl 4-oxopiperidine-1- Comercially available, carboxylate CAS:79099-07-3 2 Route 2 and N-ethyl-N-(piperidin-4- (LC/MS Method I): m/z171 [M + H]⁺ intermediates 1 yl)acetamide hydrochloride (ES⁺), at 2.21min, UV active. and 44 3 tert-butyl 2-oxo-6- Commercially available,azaspiro[3.4]octane-6- CAS: 203661-71-6 carboxylate 4 Route 1 and ethyl2-oxo-6- ¹H NMR (400 MHz, CDCl₃) δ: 1.27 (t, intermediates 3azaspiro[3.4]octane-6- J = 7.0 Hz, 3 H), 2.08 (t, J = 6.2 Hz, 2 and 5carboxylate H), 2.94-3.17 (m, 4 H), 3.49-3.59 (m, 4 H), 4.15 (q, J = 7.0Hz, 2 H) 5 ethyl carbonochloridate Commercially available, CAS: 541-41-36 benzyl 4-oxopiperidine-1- Commercially available, carboxylate CAS:19099-93-5 7 cyclopropanamine Commercially available, CAS: 765-30-0 8Route 3 and N-cyclopropyl-N-(piperidin-4- (LC/MS Method F) m/z 183 (M +H)⁺ intermediates 6 yl)acetamide (ES+) at 0.26 min, UV active and 7 9cyclobutanamine Commercially available, CAS: 2516-34-9 10 Route 2 andN-cyclobutyl-N-(piperidin-4- (LC/MS Method K) m/z 197 (M + H)⁺intermediates 1 yl)acetamide hydrochloride (ES⁺) at 3.58 min, UV activeand 9 11 Route 4 and tert-butyl 4- (LC/MS Method I) m/z 241 (M + H)⁺intermediates 1 (cyclopropylamino)piperidine- (ES+) at 4.16 min, UVactive. and 7 1-carboxylate 12 methyl bromoacetate Commerciallyavailable, CAS: 96-32-2 13 Route 4 and methyl [cyclopropyl(piperidin-(LC/MS Method K) m/z 213 (M + H)⁺ intermediates 4-yl)amino]acetate (ES⁺)at 3.17 min, UV active 11 and 12 hydrochloride 143-(chloromethyl)-1,2-oxazole Commercially available, CAS: 57684-71-6 15Route 5 and N-cyclopropyl-N-(1,2-oxazol-3- (LC/MS Method K) m/z 222 (M +H)⁺ intermediates ylmethyl)piperidin-4-amine (ES+) at 3.24 min, UVactive 11 and 14 hydrochloride 16 tert-butyl 6-oxo-2- Commerciallyavailable, azaspiro[3.3]heptane-2- CAS: 1181816-12-5 carboxylate 17Route 1 and ethyl 6-oxo-2- ¹H NMR (400 MHz, CDCl₃) δ: 1.26 (t,Intermediates 5 azaspiro[3.3]heptane-2- J = 6.6 Hz, 3 H), 3.31 (s, 4 H),and 16 carboxylate 4.06-4.24 (m, 6 H) 18 Route 6 and N-ethyl-N-(2,2,2-(LC/MS Method F) m/z 211 (M + H)⁺ intermediatestrifluoroethyl)piperidin-4-amine (ES+) at 1.41 min, UV active 1, 19 and20 hydrochloride 19 2,2,2-trifluoroethanamine Commercially available,CAS: 753-90-2 20 acetaldehyde Commercially available, CAS: 75-07-0 21Route 7 and N-cyclopropyl-N-(2- (LC/MS Method K) m/z 199 (M + H)⁺intermediates methoxyethyl)piperidin-4- (ES+) at 3.64 min, UV active 11and 22 amine hydrochloride 22 1-bromo-2-methoxyethane Commerciallyavailable, CAS: 6482-24-2 23 Route 2 and N-cyclobutyl-N-(piperidin-4-(LC/MS Method K) m/z 197 (M + H)⁺ intermediates 1 yl)acetamidehydrochloride (ES+) at 3.58 min, UV active and 9 24 Route 2 andN-methyl-N-(piperidin-4- (LC/MS Method K) m/z 157 (M + H)⁺ intermediates1 yl)acetamide hydrochloride (ES+) at 3.21 min, UV active and 25 25methylamine Commercially available, CAS: 74-89-5 26 Route 2 andN-(piperidin-4-yl)-N-(propan-2- (LC/MS Method K) m/z 185 (M + H)⁺intermediates 1 yl)acetamide hydrochloride (ES+) at 3.31 min, UV activeand 27 27 isopropylamine Commercially available, CAS: 75-31-0 28 Route 2and N-cyclopropyl-N-(piperidin-4- (LC/MS Method K) m/z 197 (M + H)⁺intermediates yl)propanamide hydrochloride (ES+) at 3.21 min, UV active1, 7 and 29 29 propanoyl chloride Commercially available, CAS: 79-03-830 Route 1 and ethyl 6-oxo-2- ¹H NMR (400 MHz, MeOD-d₄) δ: Intermediates5 azaspiro[3.4]octane-2- 1.24 (q, J = 7.0 Hz, 3 H), 2.16-2.32 (m, 4 and31 carboxylate H), 2.47 (s, 2 H), 3.85-3.97 (m, 4 H), 4.08 (q, J = 7.0Hz, 2 H) 31 tert-butyl 6-oxo-2- Commercially available,azaspiro[3.4]octane-2- CAS: 1363382-39-1 carboxylate 32 Route 2 andN-cyclopropyl-2,2,2-trifluoro-N- (LC/MS Method K) m/z 237 (M + H)⁺intermediates (piperidin-4-yl)acetamide (ES+) at 3.66 min, UV active 1,7 and 33 hydrochloride 33 trifluoroacetic anhydride Commerciallyavailable, CAS: 407-25-0 34 tert-butyl 4-formylpiperidine-1-Commercially available, carboxylate CAS: 137076-22-3 35 Route 8 andN-cyclopropyl-N-(piperidin-4- (LC/MS Method C): m/z 197 (M + H)⁺Intermediates 7 ylmethyl)acetamide (ES⁺), at 0.87 min, UV active and 34hydrochloride 36 Route 9 and N-cyclopropyl-N-(piperidin-4- (LC/MS MethodC): m/z 211 (M + H)⁺ intermediates ylmethyl)propanamide (ES⁺), at 1.06min, UV active. 7, 29 and 34 hydrochloride 37 tert-butyl4-acetylpiperidine-1- Commercially available, carboxylate CAS:206989-61-9 38 Route 8 and N-cyclopropyl-N-[1-(piperidin- (LC/MS MethodC): m/z 211 (M + H)⁺ intermediates 7 4-yl)ethyl]acetamide (ES⁺), at 1.07min, UV active and 37 hydrochloride 39 Route 9 andN-cyclopropyl-N-[1-(piperidin- (LC/MS Method C): m/z 225 (M + H)⁺intermediates 4-yl)ethyl]propanamide (ES⁺), at 1.34 min, UV active. 7,29 and 37 hydrochloride 40 methyl carbonochloridate Commerciallyavailable, CAS: 79-22-1 41 Route 1 and methyl 2-oxo-6- ¹H NMR: (400 MHz,MeOD-d₄) δ: intermediates 3 azaspiro[3.4]octane-6- 2.06-2.15 (m, 2 H),2.94-3.04 (m, 2 and 40 carboxylate H), 3.05-3.17 (m, 2 H), 3.47 (td, J =6.8, 2.5 Hz, 2 H), 3.54 (d, J = 2.5 Hz, 2 H), 3.69 (s, 3 H) 42 Route 10N-ethyl-N-(piperidin-4-yl) (LC/MS Method K): m/z 157 (M + H)⁺ andformamide (ES⁺), at 2.31 min, UV active. intermediate 52 43 Route 2 andtert-butyl 4- (LC/MS Method F): m/z 229 [M + H]⁺ intermediates 1(ethylamino)piperidine-1- (ES⁺), at 1.52 min, UV active. and 44carboxylate 44 ethanamine Commercially available, CAS: 75-04-7 45propan-1-amine Commercially available, CAS: 107-10-8 46 Route 11 andN-(piperidin-4-yl)-N- (LC/MS Method K): m/z 185 (M + H)⁺ intermediates 1propylacetamide (ES⁺), at 3.13 min, UV active. and 45 trifluoroacetate47 Route 11 and N-(2-methylpropyl)-N- (LC/MS Method I): m/z 199 (M + H)⁺intermediates 1 (piperidin-4-yl)acetamide (ES⁺), at 2.92 min, UV active.and 48 trifluoroacetate 48 2-methylpropan-1-amine Commerciallyavailable, CAS: 78-81-9 49 1-cyclopropylmethanamine Commerciallyavailable, CAS: 2516-47-4 50 Route 11 and N-(cyclopropylmethyl)-N-(LC/MS Method H): m/z 197 (M + H)⁺ intermediates 1(piperidin-4-yl)acetamide (ES⁺), at 2.68 min, UV active. and 49trifluoroacetate 51 1-(1- Commercially available,methylcyclobutyl)methanamine CAS: 1245647-53-3 52 benzyl 4- Commerciallyavailable, (ethylamino)piperidine-1- CAS: 159874-38-1 carboxylate 53Route 11 and N-ethyl-N-(piperidin-4- (LC/MS Method K): m/z 171 (M + H)⁺intermediates 1 yl)acetamide trifluoroacetate (ES⁺), at 2.24 min, UVactive. and 44 54 Route 11 and N-[(1-methylcyclobutyl)methyl]- (LC/MSMethod K): m/z 225 (M + H)⁺ intermediates 1 N-(piperidin-4-yl)acetamide(ES⁺), at 1.52 min, UV active. and 51 trifluoroacetate 551-methylcyclobutanamine Commercially available, hydrochloride CAS:174886-05-6 56 Route 11 and N-(1-methylcyclobutyl)-N- (LC/MS Method I):m/z 211 (M + H)⁺ intermediates 1 (piperidin-4-yl)acetamide (ES⁺), at3.50 min, UV active. and 55 trifluoroacetate 57 1,4-dioxa-8-Commercially available, azaspiro[4.5]decane CAS: 177-11-7 581-(3-methyloxetan-3- Commercially available, yl)methanamine CAS:153209-97-3 59 2,2-difluoroethanamine Commercially available, CAS:430-67-1 60 Route 2 and N-(2,2-difluoroethyl)-N- (LC/MS Method F): m/z207 (M + H)⁺ intermediates 1 (piperidin-4-yl)acetamide (ES⁺), at 0.39min, UV active. and 59 hydrochloride 61 2,2,2-trifluoroethanamineCommercially available, CAS: 753-90-2 62 Route 2 andN-(piperidin-4-yl)-N-(2,2,2- (LC/MS Method K): m/z 225 (M + H)⁺intermediates 1 trifluoroethyl)acetamide (ES⁺), at 3.21 min, UV active.and 61 hydrochloride 63 2-fluoro-2-methylpropan-1- Commerciallyavailable, amine CAS: 879121-42-3 64 Route 2 andN-(2-fluoro-2-methylpropyl)-N- (LC/MS Method I): m/z 217 (M + H)⁺intermediates 1 (piperidin-4-yl)acetamide (ES⁺), at 2.96 min, UV active.and 63 hydrochloride 65 2,2-difluoropropan-1-amine Commerciallyavailable, CAS: 421-00-1 66 Route 11 and N-(2,2-difluoropropyl)-N-(LC/MS Method I): m/z 221 (M + H)⁺ intermediates 1(piperidin-4-yl)acetamide (ES⁺), at 2.82 min, UV active. and 65trifluoroacetate 67 3,3-difluorocyclobutanamine Commercially available,CAS: 791061-00-2 68 Route 11 and N-(3,3-difluorocyclobutyl)-N- (LC/MSMethod I): m/z 233 (M + H)⁺ intermediates 1 (piperidin-4-yl)acetamide(ES⁺), at 3.02 min, UV active. and 67 trifluoroacetate 692-methoxy-2-methylpropan-1- Commercially available, amine CAS:89282-70-2 70 Route 2 and N-(2-methoxy-2-methylpropyl)-N- (LC/MS MethodI): m/z 229 (M + H)⁺ intermediates 1 (piperidin-4-yl)acetamide (ES⁺), at2.97 min, UV active. and 69 hydrochloride 71 Route 11 and N-[(1- (LC/MSMethod F): m/z 241 (M + H)⁺ intermediates 1 methoxycyclobutyl)methyl]-N-(ES⁺), at 1.29 min, UV active. and 72 (piperidin-4-yl)acetamidetrifluoroacetate 72 1-(1- Commercially available,methoxycyclobutyl)methanamine CAS: 1443980-50-4 731-(aminomethyl)cyclobutanol Commercially available, CAS: 180205-28-1 74Route 11 and N-[(1- (LC/MS Method J): m/z 227 (M + H)⁺ intermediates 1hydroxycyclobutyl)methyl]-N- (ES⁺), at 2.97 min, UV active. and 73(piperidin-4-yl)acetamide trifluoroacetate 75 O-methylhydroxylamineCommercially available, CAS: 67-62-9 76 Route 12 andN-methoxy-N-(piperidin-4- (LC/MS Method F): m/z 173 (M + H)⁺intermediates 1 yl)acetamide trifluoroacetate (ES⁺), at 0.25 min, UVactive. and 75 77 aniline Commercially available, CAS: 62-53-3 78 Route2 and N-phenyl-N-(piperidin-4-yl) (LC/MS Method I): m/z 219 (M + H)⁺intermediates 1 acetamide hydrochloride (ES⁺), at 3.18 min, UV active.and 77 79 2-Aminopyridine Commercially available, CAS: 504-29-0 80 Route2 and N-(piperidin-4-yl)-N-(pyridin-2- (LC/MS Method I): m/z 220 (M +H)⁺ intermediates 1 yl) acetamide hydrochloride (ES⁺), at 2.41 min, UVactive. and 79 81 benzylamine Commercially available, CAS: 100-46-9 82Route 2 and N-benzyl-N-(piperidin-4- (LC/MS Method I): m/z 233 (M + H)⁺intermediates 1 yl)acetamide hydrochloride (ES⁺), at 3.17 min, UVactive. and 81 83 [(tert- Commercially available,butoxycarbonyl)amino]acetic CAS: 4530-20-5 acid 84 N-methoxymethanamineCommercially available, hydrochloride CAS: 6638-79-5 85N-(3-dimethylaminopropyl)-N′- Commercially available, ethylcarbodiimideCAS: 25952-53-8 hydrochloride 86 1-hydroxybenzotriazole Commerciallyavailable, hydrate CAS: 123333-53-9 87 lithium aluminium hydrideCommercially available, CAS: 16853-85-3 88 p-toluenesulfonylmethylCommercially available, isocyanide CAS: 36635-61-7 89 Route 13 and1-(1,3-oxazol-5- ¹H NMR (400 MHz, DMSO-d₆) δ: intermediatesyl)methanamine hydrochloride 4.11-4.17 (m, 2 H), 7.28 (s, 1 H), 83, 84,85, 86, 8.47 (t, J = 12.0 Hz, 1 H), 8.77 (s, 3 87 and 88 H), 10.23 (s, 1H). 90 Route 2 and N-(1,3-oxazol-5-ylmethyl)-N- (LC/MS Method I): m/z224 (M + H)⁺ intermediates 1 (piperidin-4-yl)acetamide (ES⁺), at 2.41min, UV active. and 89 hydrochloride 91 tert-butyl 4-aminopiperidine-1-Commercially available, carboxylate CAS: 87120-72-7 921,3-oxazole-2-carbaldehyde Commercially available, CAS: 65373-52-6 93Route 11 and N-(1,3-oxazol-2-ylmethyl)-N- (LC/MS Method F): m/z 224 (M +H)⁺ intermediates (piperidin-4-yl)acetamide (ES⁺), at 0.31 min, UVactive. 91 and 92 trifluoroacetate 94 Route 2 and methylethyl(piperidin-4- (LC/MS Method F): m/z 187 (M + H)⁺ intermediates 1yl)carbamate hydrochloride (ES⁺), at 0.90 min, UV active and 40 95 Route2 and ethyl ethyl(piperidin-4- (LC/MS Method F): m/z 201 (M + H)⁺intermediates 1 yl)carbamate hydrochloride (ES⁺), at 1.52 min, UV activeand 5 96 Route 11 and Methyl isopropyl(piperidin-4- (LC/MS Method I):m/z 201 (M + H)⁺ intermediates yl)carbamate trifluoroacetate (ES⁺), at2.97 min, UV active 1, 27 and 40 97 Route 6 and tert-butyl 4-[(2,2,2-(LC/MS Method I): m/z 227 (M + H − 56)⁺ intermediates 1trifluoroethyl)amino]piperidine- (ES⁺), at 4.62 min, UV active and 191-carboxylate 98 Route 14 and methyl piperidin-4-yl(2,2,2- (LC/MS MethodI): m/z 241 (M + H)⁺ intermediates trifluoroethyl)carbamate (ES⁺), at3.34 min, UV active 97 and 40 hydrochloride 99 Route 15 and2-methoxyethyl piperidin-4- (LC/MS Method I): m/z 285 (M + H)⁺intermediates yl(2,2,2- (ES⁺), at 3.38 min, UV active 97 and 22trifluoroethyl)carbamate hydrochloride 100 dimethylcarbamic chlorideCommercially available, CAS: 79-44-7 101 Route 2 and1-ethyl-3,3-dimethyl-1- (LC/MS Method F): m/z 200 (M + H)⁺ intermediates1 piperidin-4-ylurea (ES⁺), at 0.92 min, UV active and 100 hydrochloride102 2-fluoroethanamine Commercially available, hydrochloride CAS:460-08-2 103 Route 6 and N-ethyl-N-(2- (LC/MS Method F) m/z 175 (M + H)⁺intermediates fluoroethyl)piperidin-4-amine (ES+) at 0.26 min, UV active1, 102 and 20 hydrochloride 104 Difluoroethan-1-amine Commerciallyavailable, CAS: 430-67-1 105 Route 6 and N-(2,2-difluoroethyl)-N- (LC/MSMethod I) m/z 193 (M + H)⁺ intermediates ethylpiperidin-4-amine (ES+) at3.60 min, UV active 1, 104 and 20 hydrochloride 106 oxetan-3-amineCommercially available, CAS: 21635-88-1 107 N-methyl-2-pyrrolidinoneCommercially available, CAS: 872-50-4 108 2,2,2-trifluoroethylCommercially available, trifluoromethanesulfonate CAS: 6226-25-1 109Route 16 and N-cyclopropyl-N-(2,2,2- (LC/MS Method K) m/z 223 (M + H)⁺intermediates trifluoroethyl)piperidin-4-amine (ES+) at 5.08 min, UVactive 11, 107 and 108 hydrochloride 110 2-bromoethanol Commerciallyavailable, CAS: 540-51-2 111 Route 17 and 2-[cyclopropyl(piperidin-4-(LC/MS Method F) m/z 185 (M + H)⁺ intermediates yl)amino]ethanol (ES+)at 0.26 min, UV active 11 and 110 trifluoroacetate 112 Route 18 andtert-butyl 4- (LC/MS Method I) m/z 255 (M + H)⁺ intermediates 1(cyclobutylamino)piperidine-1- (ES+) at 4.38 min, UV active and 9carboxylate 113 Route 17 and 2-[cyclobutyl(piperidin-4- (LC/MS Method I)m/z 199 (M + H)⁺ intermediates yl)amino]ethanol (ES+) at 3.20 min, UVactive 112 and 110 trifluoroacetate 114 N-ethyl-N-(piperidin-4-Commercially available, yl)acetamide CAS: 139062-99-0 115 4-nitrophenylCommercially available, carbonochloridate CAS: 7693-46-1 116(1,1-²H₂)-ethanol Commercially available, CAS: 1859-09-2 117 Route 7 and2-[piperidin-4-yl(2,2,2- (LC/MS Method I): m/z 227 (M + H)⁺Intermediates trifluoroethyl)amino]ethanol (ES⁺), at 4.40 min, UVactive. 97 and 110 hydrochloride 118 2-methoxyethylamine Commerciallyavailable, CAS: 109-85-3 119 Route 19 and N-(2-methoxyethyl)-N-(2,2,2-(LC/MS Method I): m/z 241 (M + H)⁺ Intermediatestrifluoroethyl)piperidin-4-amine (ES⁺), at 4.05 min, UV active. 1, 118and 108 trifluoroacetate 120 Route 20 and N-ethyl-N-(oxetan-3- (LC/MSMethod I): m/z 185 (M + H)⁺ Intermediates yl)piperidin-4-amine (ES⁺), at2.42 min, UV active. 1, 106, 121 and trifluoroacetate 20 121 Route 20and tert-butyl 4-(oxetan-3- (LC/MS Method I): m/z 257 (M + H)⁺Intermediates 1 ylamino)piperidine-1- (ES⁺), at 2.92 min, UV active. and106 carboxylate 122 Route 19 and N-(oxetan-3-yl)-N-(2,2,2- (LC/MS MethodI): m/z 239 (M + H)⁺ Intermediates trifluoroethyl)piperidin-4-amine(ES⁺), at 3.18 min, UV active. 121 and 108 trifluoroacetate 1233-aminopropanenitrile Commercially available, CAS: 151-18-8 124 Route 20and 3-[ethyl(piperidin-4- (LC/MS Method I): m/z 182 (M + H)⁺Intermediates yl)amino]propanenitrile (ES⁺), at 3.16 min, UV active. 1,123 and 20 trifluoroacetate 125 2-(methylsulfonyl)ethanamineCommercially available, CAS: 49773-20-8 126 Route 6 and N-ethyl-N-[2-(LC/MS Method I): m/z 235 (M + H)⁺ Intermediates(methylsulfonyl)ethyl]piperidin- (ES⁺), at 2.71 min, UV active. 1, 125and 20 4-amine hydrochloride 127 iodoethane Commercially available, CAS:75-03-6 128 Route 21 and N-ethyl-N-methoxypiperidin-4- (LC/MS Method F):m/z 159 (M + H)⁺ Intermediates amine trifluoroacetate (ES⁺), at 2.71min, UV active. 1, 75 and 127 129 ethyl bromoacetate Commerciallyavailable, CAS: 105-36-2 130 Route 22 and 2-[phenyl(piperidin-4- (LC/MSMethod F): m/z 221 (M + H)⁺ Intermediates yl)amino]ethanol hydrochloride(ES⁺), at 1.09 min, UV active. 1, 77 and 129 131 Route 18 and tert-butyl4- (LC/MS Method I): m/z 291 (M + H)⁺ Intermediates 1(benzylamino)piperidine-1- (ES⁺), at 4.80 min, UV active and 81carboxylate 132 Route 16 and N-benzyl-N-(2,2,2- (LC/MS Method K): m/z273 (M + H)⁺ Intermediates trifluoroethyl)piperidin-4-amine (ES⁺), at5.99 min, UV active. 131, 107 and 11 hydrochloride 133 Route 20 andN-ethyl-N-(1,3-oxazol-5- ¹H NMR (400 MHz, DMSO-d₆) δ: Intermediatesylmethyl)piperidin-4-amine 1.02-1.16 (m, 3 H), 1.74-2.02 (m, 2 1, 89 and20 trifluoroacetate H), 2.14-2.31 (m, 2 H), 2.89-3.03 (m, 2 H),3.08-3.32 (m, 3 H), 3.37-3.48 (m, 2 H), 4.61 (s, 2 H), 7.52 (s, 1 H),8.59 (s, 1 H). Exchangeable protons not observed. 134 2-fluoroethylCommercially available, trifluoromethanesulfonate CAS: 95353-04-1 135Route 19 and N-(2-fluoroethyl)-N-(1,3- (LC/MS Method I): m/z 228 (M +H)⁺ Intermediates oxazol-5-ylmethyl)piperidin-4- (ES⁺), at 3.19 min, UVactive. 1, 89 and 134 amine trifluoroacetate 136 Route 19 andN-(1,3-oxazol-5-ylmethyl)-N- (LC/MS Method F): m/z 264 (M + H)⁺Intermediates (2,2,2-trifluoroethyl)piperidin-4- (ES⁺), at 1.53 min, UVactive. 1, 89 and 108 amine trifluoroacetate 137 tert-butyl 4-Commercially available, propanoylpiperidine-1- CAS: 419571-73-6carboxylate 138 Route 23 and N-ethyl-N-[1-(piperidin-4- (LC/MS MethodI): m/z 213 (M + H)⁺ Intermediates yl)propyl]acetamide (ES⁺), at 3.12min, UV active. 44 and 137 trifluoroacetate 139 tert-butyl4-(2-aminopropan-2- Commercially available, yl)piperidine-1-carboxylateCAS: 530116-33-7 140 Route 24 and N-ethyl-N-[2-(piperidin-4- (LC/MSMethod F): m/z 213 (M + H)⁺ Intermediates yl)propan-2-yl]acetamide(ES⁺), at 1.50 min, UV active. 139 and 20 hydrochloride 141 tert-butyl4-(1- Commercially available, hydroxyethyl)piperidine-1- CAS:183170-69-6 carboxylate 142 methanesulfonyl chloride Commerciallyavailable, CAS: 124-63-0 143 1H-pyrazole Commercially available, CAS:288-13-1 144 Route 25 and 4-[1-(1H-pyrazol-1- (LC/MS Method I): m/z 180(M + H)⁺ Intermediates yl)ethyl]piperidine (ES⁺), at 3.42 min, UVactive. 141, 142 and trifluoroacetate 143 145 tert-butyl4-hydroxypiperidine- Commercially available, 1-carboxylate CAS:109384-19-2 146 (1-bromoethyl)benzene Commercially available, CAS:585-71-7 147 Route 26 and 4-(1-phenylethoxy)piperidine (LC/MS Method I):m/z 206 (M + H)⁺ Intermediates hydrochloride (ES⁺), at 4.93 min, UVactive. 145 and 146 148 tert-butyl 4-bromopiperidine-1- Commerciallyavailable, carboxylate CAS: 180695-79-8 149 phenylmethanethiolCommercially available, CAS: 100-53-8 150 Route 27 and4-(benzylsulfanyl)piperidine (LC/MS Method F): m/z 208 (M + H)⁺Intermediates hydrochloride (ES⁺), at 1.70 min, UV active. 148 and 149151 tert-butyl 6-oxo-2- Commercially available, azaspiro[3.4]octane-2-CAS: 1363382-39-1 carboxylate 152 Route 28 and methyl 6-oxo-2- (LC/MSMethod I): m/z 184 (M + H)⁺ Intermediates azaspiro[3.4]octane-2- (ES⁺),at 2.47 min, UV active. 151 and 40 carboxylate 153 Route 28 and ethyl6-oxo-2- (LC/MS Method I): m/z 198 (M + H)⁺ Intermediatesazaspiro[3.4]octane-2- (ES⁺), at 2.89 min, UV active. 151 and 5carboxylate

TABLE 3 LCMS Ex. Inter- Syn- Meth- No. Name mediate thetic ¹ HNMR odLCMS data 1-1 Ethyl 6-{4- 8 and 17 a (400 MHz, CDCl₃) δ: 0.80-0.94 (m, 4H), 1.25 (t, J = 7.0 I m/z 350 (M + H)⁺ [acetyl(cyclopropyl)amino] Hz, 3H), 1.73-2.07 (m, 8 H), 2.22 (s, 3 H), 2.30-2.35 (ES⁺) atpiperidin-1-yl}-2- (m, 2 H), 2.46-2.62 (m, 2 H), 2.88-2.91 (m, 2 H),3.88 3.45 min, UV active azaspiro[3.3]heptane-2- (s, 2 H), 4.00 (s, 2H), 4.08-4.20 (m, 3 H) carboxylate 1-2 Ethyl 6-{4- 2 and 17 a (400 MHz,MeOD-d₄) δ:1.10-1.30 (m, 6 H), 1.62-1.73 I m/z 338 (M + H)⁺[acetyl(ethyl)amino]piperi (m, 1 H), 1.73-2.00 (m, 5 H), 2.02-2.11 (m, 2H), 2.14 (ES⁺) at din-1-yl}-2- (d, J = 2.8 Hz, 3 H), 2.34-2.44 (m, 2 H),2.63-2.76 (m, 1 3.31 min, UV active azaspiro[3.3]heptane-2- H),2.93-3.03 (m, 2 H), 3.28-3.42 (m, 2 H), 3.65-3.77 carboxylate (m, 0.5H), 3.86-3.94 (m, 2 H), 3.98-4.05 (m, 2 H), 4.09 (q, J = 7.3 Hz, 2 H),4.19-4.31 (m, 0.5 H) 2-1 Isomer 2: Ethyl 2-{4- 2 and 4 a (400 MHz,MeOD-d₄) δ:1.23-1.31 (m, 7 H), 1.68-1.99 I m/z 352 (M + H)⁺[acetyl(ethyl)amino]piperi (m, 10 H), 2.13-2.17 (m, 5 H), 2.79-3.07 (m,3 H), 3.29 (ES⁺) at din-1-yl}-6- (m, 2 H), 3.37-3.44 (m, 4 H), 4.11 (q,J = 7.0 Hz, 2 H) 3.46 min, UV active azaspiro[3.4]octane-6- carboxylate2-2 Isomer 2: Ethyl 2-{4- 4 and 26 a (400 MHz, CDCl₃) δ: 1.14-1.42 (m, 8H), 1.65-2.19 (m, I m/z 366 (M + H)⁺ [acetyl(propan-2- 18 H), 2.41-3.14(m, 3 H), 3.21-3.58 (m, 4 H), 3.85- (ES⁺) at yl)amino]piperidin-1-yl}-6-4.18 (m, 2 H) 3.99 min, UV active azaspiro[3.4]octane-6- carboxylate 2-3Isomer 1: Ethyl 2-{4- 4 and 8 a (400 MHz, DMSO-d₆) δ: 0.81-0.87 (m, 4H), 1.16-1.24 F m/z 364 (M + H)⁺ [acetyl(cyclopropyl)amino] (m, 4 H),1.80-1.91 (m, 4 H), 2.11 (s, 3 H), 2.16-2.41 (ES⁺) at piperidin-1-yl}-6-(m, 5 H), 2.64-2.90 (m, 4 H), 3.04-3.31 (m, 5 H), 3.40- 1.51 min, UVactive azaspiro[3.4]octane-6- 3.69 (m, 2 H), 4.02 (q, J = 7.0 Hz, 2 H)carboxylate 2-3 Isomer 2: Ethyl 2-{4- 4 and 8 a (400 MHz, DMSO-d₆) δ:0.67-0.90 (m, 5 H), 1.16 (t, J = F m/z 364 (M + H)⁺[acetyl(cyclopropyl)amino] 7.0 Hz, 3 H), 1.51-2.05 (m, 9 H), 2.09 (s, 3H), 2.56- (ES⁺) at piperidin-1-yl}-6- 2.68 (m, 2 H), 2.78-2.99 (m, 2 H),3.11-3.30 (m, 5 H), 1.51 min, UV active azaspiro[3.4]octane-6- 3.41-3.53(m, 1 H), 3.77-3.89 (m, 1 H), 4.00 (q, J = 7.0 carboxylate Hz, 2 H) 2-4Isomer 2: Ethyl 2-{4- 4 and 32 a (400 MHz, MeOD-d₄) δ: 0.98 (d, J = 6.5Hz, 4 H), 1.27 I m/z 418 (M + H)⁺ [cyclopropyl(trifluoroacetyl) (t, J =7.0 Hz, 3 H), 1.80-1.98 (m, 8 H), 2.11-2.27 (m, 4 H), (ES⁺) atamino]piperidin-1-yl}-6- 2.76-3.02 (m, 4 H), 3.29 (s, 2 H), 3.41 (q, J =7.0 Hz, 2 4.80 min, UV active azaspiro[3.4]octane-6- H), 3.77-3.90 (m, 1H), 4.11 (q, J = 7.0 Hz, 2 H) carboxylate 2-5 Isomer 2: Ethyl 2-{4- 4and 28 a (400 MHz, MeOD-d₄) δ: 0.80-1.00 (m, 4 H), 1.12 (t, J = I m/z378 (M + H)⁺ [cyclopropyl(propanoyl)a 7.5 Hz, 3 H), 1.25-1.36 (m, 4 H),1.73-1.76 (m, 2 H), (ES⁺) at mino]piperidin-1-yl}-6- 1.87-1.98 (m, 6 H),2.07-2.18 (m, 3 H), 2.59-2.70 (m, 2 4.10 min, UV activeazaspiro[3.4]octane-6- H), 2.77-2.84 (m, 1 H), 2.97-3.00 (m, 2 H), 3.29(s, 2 H), carboxylate 3.37-3.43 (m, 3 H), 3.91-4.02 (m, 1 H), 4.11 (q, J= 7.0 Hz, 2 H) 2-6 Isomer 2: Ethyl 2-{4- 10 and 4 a (400 MHz, DMSO-d₆)δ: 1.16 (t, J = 7.0 Hz, 3 H), 1.24- I m/z 378 (M + H)⁺[acetyl(cyclobutyl)amino] 1.40 (m, 1 H), 1.44-2.24 (m, 20 H), 2.31-2.47(m, 1 H), (ES⁺) at piperidin-1-yl}-6- 2.58-2.97 (m, 4 H), 3.15 (d, J =6.5 Hz, 2 H), 3.25-3.30 4.28 min, UV active azaspiro[3.4]octane-6- (m, 2H), 4.00 (q, J = 7.0 Hz, 2 H) carboxylate 2-7 Isomer 2: Ethyl 2-{4- 18and 4 a (400 MHz, DMSO-d₆) δ: 0.96 (t, J = 7.0 Hz, 3 H), 1.56 (t, I m/z392 (M + H)⁺ [ethyl(2,2,2- J = 7.0 Hz, 3 H), 1.31-1.40 (m, 2 H),1.61-1.99 (m, 8 H), (ES⁺) at trifluoroethyl)amino]piperi 1.94-1.99 (m, 2H), 2.58-2.68 (m, 4 H), 2.78-2.81 (m, 2 5.25 min, UV active din-1-yl}-6-H), 3.13-3.19 (m, 4 H), 3.26 (q, J = 6.5 Hz, 2 H), 3.99 (q,azaspiro[3.4]octane-6- J = 7.0 Hz, 2 H) carboxylate 2-8 Isomer 2: Ethyl2-{4- 21 and 4 a (400 MHz, MeOD-d₄) δ: 0.41-0.60 (m, 4 H), 1.27 (td, J =I m/z 380 (M + H)⁺ [cyclopropyl(2- 7.0, 2.5 Hz, 3 H), 1.64-1.73 (m, 2H), 1.86-2.00 (m, 9 (ES⁺) at methoxyethyl)amino]pipe H), 2.14-2.18 (m, 2H), 2.68-2.89 (m, 4 H), 3.01-3.04 4.38 min, UV active ridin-1-yl}-6- (m,2 H), 3.34 (s, 3 H), 3.37-3.40 (m, 4 H), 3.53 (t, J = 6.0azaspiro[3.4]octane-6- Hz, 2 H), 4.12 (q, J = 7.0 Hz, 2 H) carboxylate2-9 Isomer 2: Ethyl 2-{4- 13 and 4 a (400 MHz, DMSO-d₆) δ: 0.28-0.34 (m,2 H), 0.40-0.47 I m/z 394 (M + H)⁺ [cyclopropyl(2-methoxy- (m, 2 H),1.16 (t, J = 7.0 Hz, 3 H), 1.32-1.41 (m, 2 H), (ES⁺) at 2- 1.62 (t, J =11.5 Hz, 2 H), 1.72-1.85 (m, 6 H), 1.94-1.98 4.35 min, UV activeoxoethyl)amino]piperidin- (m, 2 H), 2.17-2.23 (m, 1 H), 2.55-2.70 (m, 2H), 2.76- 1-yl}-6- 2.79 (m, 2 H), 3.14 (d, J = 6.5 Hz, 2 H), 3.24-3.29(m, 2 azaspiro[3.4]octane-6- H), 3.43 (s, 2 H), 3.59 (s, 3 H), 3.99 (q,J = 7.0 Hz, 2 H) carboxylate 2-10 Isomer 2: Ethyl 2-{4- 15 and 4 a (400MHz, CDCl₃) δ: 0.35-0.44 (m, 2 H), 0.48-0.60 (m, I m/z 403 (M + H)⁺[cyclopropyl(1,2-oxazol- 2 H), 1.26 (t, J = 7.0 Hz, 3 H), 1.58-2.14 (m,14 H), 2.49- (ES⁺) at 3- 3.08- (m, 3H), 3.24-3.52 (m, 4 H), 3.95 (s, 2H), 4.12 4.43 min, UV active ylmethyl)amino]piperidin- (q, J = 7.0 Hz, 2H), 6.30 (s, 1 H), 8.34 (s, 1 H) 1-yl}-6-3.08 (m, 3 H), 3.24azaspiro[3.4]octane-6- carboxylate 2-11 Isomer 2: Ethyl 2-(4- 4 and 35 b(400 MHz, CDCl₃) δ: 0.66-0.75 (m, 2 H), 0.80-0.93 (m, E m/z 378 (M + H)⁺{[acetyl(cyclopropyl)amino] 2 H), 1.21-1.41 (m, 5 H), 1.59-2.10 (m, 11H), 2.20 (s, 3 (ES⁺) at methyl}piperidin-1-yl)-6- H), 2.59-2.76 (m, 2H), 2.79-2.94 (m, 2 H), 3.19-3.44 3.92 min, UV activeazaspiro[3.4]octane-6- (m, 6 H), 4.09 (q, J = 7.0 Hz, 2 H) carboxylate2-12 Isomer 2: Ethyl 2-(4- 4 and 36 b (400 MHz, CDCl₃) δ: 0.64-0.73 (m,2 H), 0.83-0.91 (m, E m/z 392 (M + H)⁺ {[cyclopropyl(propanoyl) 2 H),1.12 (t, J = 7.5 Hz, 3 H), 1.21-1.40 (m, 5 H), 1.57- (ES⁺) atamino]methyl}piperidin-1- 2.12 (m, 11 H), 2.45-2.72 (m, 4 H), 2.74-2.93(m, 2 H), 4.28 min, UV active yl)-6-azaspiro[3.4]octane- 3.17-3.44 (m, 6H), 4.09 (q, J = 7.0 Hz, 2 H) 6-carboxylate 2-13 Isomer 2: Ethyl2-(4-{1- 4 and 38 b (400 MHz, CDCl₃) δ: 0.70-0.89 (m, 4 H), 1.08-1.31(m, E m/z 392 (M + H)⁺ [acetyl(cyclopropyl)amino] 8 H), 1.46-2.09 (m, 11H), 2.16 (s, 3 H), 2.54-2.72 (m, 2 (ES⁺) at ethyl}piperidin-1-yl)-6- H),2.78-2.95 (m, 2 H), 3.23-3.41 (m, 5 H), 4.09 (q, J = 4.16 min, UV activeazaspiro[3.4]octane-6- 7.0 Hz, 2 H) carboxylate 2-14 Isomer 2: Ethyl2-(4-{1- 4 and 39 b (400 MHz, CDCl₃) δ: 0.67-0.78 (m, 2 H), 0.79-0.94(m, E m/z 406 (M + H)⁺ [cyclopropyl(propanoyl) 2 H), 1.01-1.34 (m, 11H), 1.50 (d, J = 11.5 Hz, 1 H), (ES⁺) at amino]ethyl}piperidin-1-1.57-2.15 (m, 10 H), 2.39-2.73 (m, 4 H), 2.75-2.98 (m, 4.52 min, UVactive yl)-6-azaspiro[3.4]octane-6- 2 H), 3.16-3.47 (m, 5 H), 4.09 (d, J= 7.0 Hz, 2 H). carboxylate 2-15 Isomer 2: Methyl 2-{4- 4 and 23 a (400MHz, DMSO-d₆) δ: 1.22-1.41 (m, 1 H), 1.46-2.23 I m/z 364 (M + H)⁺[acetyl(cyclobutyl)amino] (m, 18 H), 2.35-2.45 (m, 1 H), 2.58-2.70 (m, 2H), 2.73- (ES⁺) at piperidin-1-yl}-6- 2.92 (m, 3 H), 3.15 (d, J = 3.5Hz, 2 H), 3.26-3.31 (m, 3 3.99 min, UV active azaspiro[3.4]octane-6- H),3.56 (s, 3 H) carboxylate 2-16 Isomer 2: Ethyl 2-{4- 4 and 42 a (400MHz, MeOD-d₄) δ: 1.17 (t, J = 6.8 Hz, 2 H), 1.23- I m/z 338 (M + H)⁺[ethyl(formyl)amino]piperi 1.44 (m, 4 H), 1.56-1.95 (m, 9 H), 2.03-2.28(m, 2 H), (ES⁺) at din-1-yl}-6- 2.67-2.92 (m, 1 H), 3.01 (d, J = 9.5 Hz,2 H), 3.29 (s, 2 3.50 min, UV active azaspiro[3.4]octane-6- H),3.38-3.46 (m, 4 H), 3.46-3.66 (m, 1 H), 4.11 (q, J = carboxylate 6.8 Hz,2 H), 4.66 (s, 1 H), 8.06-8.18 (m, 1 H) 2-17 Isomer 1: Methyl 2-{4- 41and 53 a (400 MHz, DMSO-d₆) δ: 0.99 (t, J = 6.9 Hz, 1 H), 1.09 (t, I m/z338 (M + H)⁺ [acetyl(ethyl)amino]piperi J = 7.2 Hz, 2 H), 1.43-1.49 (m,1 H), 1.57-1.83 (m, 9 H), (ES⁺) at din-1-yl}-6- 1.94-2.05 (m, 5 H),2.57-2.66 (m, 2 H), 2.82 (d, J = 9.8 3.17 min, UV activeazaspiro[3.4]octane-6- Hz, 2 H), 3.12-3.29 (m, 6 H), 3.57 (s, 3 H)carboxylate 2-17 Isomer 2: Methyl 2-{4- 41 and 53 a (400 MHz, DMSO-d₆)δ: 0.98 (t, J = 6.7 Hz, 1 H), 1.08 (t, I m/z 338 (M + H)⁺[acetyl(ethyl)amino]piperi J = 7.0 Hz, 2 H), 1.42-1.51 (m, 1 H),1.51-1.90 (m, 9 H), (ES⁺) at din-1-yl}-6- 1.91-2.04 (m, 5 H), 2.59-2.65(m, 2 H), 2.80 (d, J = 10.4 3.24 min, UV active azaspiro[3.4]octane-6-Hz, 2 H), 3.08-3.32 (m, 6 H), 3.54 (s, 3 H) carboxylate 2-18 Isomer 2:Ethyl 2-{4- 4 and 46 a (400 MHz, DMSO-d₆) δ: 0.73-0.92 (m, 3 H), 1.16(t, J = I m/z 366 (M + H)⁺ [acetyl(propyl)amino]pipe 7.0 Hz, 3 H),1.35-1.54 (m, 3 H), 1.56-1.77 (m, 7 H), (ES⁺) at ridin-1-yl}-6-1.77-1.88 (m, 2 H), 1.92-2.04 (m, 5 H), 2.59-2.66 (m, 1 3.82 min, UVactive azaspiro[3.4]octane-6- H), 2.76-2.86 (m, 2 H), 2.99-3.12 (m, 2H), 3.14 (d, J = carboxylate 5.1 Hz, 2 H), 3.28 (d, J = 6.6 Hz, 2 H),3.42-3.57 (m, 1 H), 4.00 (q, J = 7.1 Hz, 2 H) 2-19 Isomer 1: Ethyl 2-{4-4 and 47 a (400 MHz, MeOD-d₄) δ: 0.90 (d, J = 6.8 Hz, 3 H), 0.98 (d, Im/z 380 (M + H)⁺ [acetyl(2- J = 6.8 Hz, 3 H), 1.17-1.34 (m, 3 H),1.65-1.86 (m, 3 H), (ES⁺) at methylpropyl)amino]piper 1.86-1.96 (m, 6H), 1.97-2.08 (m, 2 H), 2.10-2.19 (m, 5 4.06 min, UV activeidin-1-yl}-6- H), 2.79 (t, J = 7.3 Hz, 1 H), 2.93-3.42 (m, 2 H), 3.17(dd, azaspiro[3.4]octane-6- J = 11.7, 7.8 Hz, 2 H), 3.33 (s, 2 H),3.37-3.42 (m, 2 H), carboxylate 3.61-3.95 (m, 1 H), 4.12 (q, J = 7.2 Hz,2 H) 2-19 Isomer 2: Ethyl 2-{4- 4 and 47 a (400 MHz, MeOD-d₄) δ: 0.90(d, J = 6.8 Hz, 3 H), 0.98 (d, I m/z 380 (M + H)⁺ [acetyl(2- J = 6.4 Hz,3 H), 1.27 (t, J = 6.8 Hz, 3 H), 1.64-1.83 (m, 3 (ES⁺) atmethylpropyl)amino]piper H), 1.85-2.14 (m, 11 H), 2.14-2.30 (m, 2 H),2.80 (dq, J 4.20 min, UV active idin-1-yl}-6- = 14.8, 7.6 Hz, 1 H),2.92-3.06 (m, 2 H), 3.17 (dd, J = azaspiro[3.4]octane-6- 11.7, 7.8 Hz, 2H), 3.28 (s, 2 H), 3.40 (d, J = 6.4 Hz, 2 H), carboxylate 3.64-3.96 (m,1 H), 4.11 (q, J = 7.2 Hz, 2 H) 2-20 Isomer 1: Ethyl 2-{4- 4 and 50 a(400 MHz, MeOD-d₄) δ: 1.38-1.56 (m, 2 H), 1.58-1.70 I m/z 378 (M + H)⁺[acetyl(cyclopropylmethyl) (m, 1 H), 1.72-1.88 (m, 1 H), 2.02-2.30 (m, 1H), 2.32- (ES⁺) at amino]piperidin-1-yl}-6- 2.51 (m, 4 H), 2.85 (d, J =11.8 Hz, 1 H), 2.89-3.19 (m, 9 3.79 min, UV activeazaspiro[3.4]octane-6- H), 3.22-3.29 (m, 2 H), 3.80-4.01 (m, 1 H),4.03-4.21 carboxylate (m, 3 H), 4.36 (dd, J = 17.4, 6.4 Hz, 2 H),4.43-4.63 (m, 5 H), 4.75-4.95 (m, 1 H), 5.26 (q, J = 7.0 Hz, 2 H) 2-20Isomer 2: Ethyl 2-{4- 4 and 50 a (400 MHz, MeOD-d₄) δ: 1.48 (dd, J =13.2, 5.1 Hz, 2 H), I m/z 378 (M + H)⁺ [acetyl(cyclopropylmethyl) 1.63(d, J = 7.3 Hz, 1 H), 1.78 (d, J = 7.5 Hz, 1 H), 2.04- (ES⁺) atamino]piperidin-1-yl}-6- 2.26 (m, 1 H), 2.41 (t, J = 7.0 Hz, 4 H),2.76-2.90 (m, 1 3.90 min, UV active azaspiro[3.4]octane-6- H), 2.90-3.05(m, 3 H), 3.05-3.19 (m, 7 H), 3.28 (s., 2 carboxylate H), 3.87-4.04 (m,1 H), 4.06-4.22 (m, 2 H), 4.34 (d, J = 6.5 Hz, 1 H), 4.38 (d, J = 6.0Hz, 1 H), 4.42 (s, 2 H), 4.45- 4.48 (m, 1 H), 4.55 (q, J = 6.4 Hz, 2 H),4.76-4.97 (m, 1 H), 5.25 (q, J = 7.0 Hz, 2 H) 2-21 Isomer 1: Ethyl 2-(4-4 and 54 a (400 MHz, MeOD-d₄) δ: 1.13 (s, 1 H), 1.20-1.31 (m, 5 H), Im/z 406 (M + H)⁺ {acetyl[(1- 1.56-1.67 (m, 2 H), 1.67-1.97 (m, 10 H),1.98-2.28 (m, (ES⁺) at methylcyclobutyl)methyl] 8 H), 2.45 (q, J = 12.3Hz, 1 H), 2.64-2.85 (m, 1 H), 2.97 4.59 min, UV activeamino}piperidin-1-yl)-6- (t, J = 9.5 Hz, 2 H), 3.26 (s, 2 H), 3.33-3.43(m, 4 H), azaspiro[3.4]octane-6- 3.59-3.76 (m, 1 H), 4.11 (q, J = 7.0Hz, 2 H) carboxylate 2-21 Isomer 2: Ethyl 2-(4- 4 and 54 a (400 MHz,MeOD-d₄) δ: 1.12 (s, 1 H), 1.18-1.37 (m, 5 H), I m/z 406 (M + H)⁺{acetyl[(1- 1.45 (s, 1 H), 1.59-1.89 (m, 7 H), 1.89-1.98 (m, 4 H), (ES⁺)at methylcyclobutyl)methyl] 1.98-2.23 (m, 8 H), 2.31-2.56 (m, 1 H),2.69-2.86 (m, 1 4.71 min, UV active amino}piperidin-1-yl)-6- H), 2.98(d, J = 10.1 Hz, 2 H), 3.26 (s, 2 H), 3.33-3.44 azaspiro[3.4]octane-6-(m, 4 H), 3.55-3.79 (m, 1 H), 4.09 (q, J = 7.0 Hz, 2 H) carboxylate 2-22Isomer 2: Ethyl 2-{4- 4 and 56 a (400 MHz, MeOD-d₄) δ: 1.25 (t, J = 7.0Hz, 3 H), 1.46- I m/z 392 (M + H)⁺ [acetyl(1- 1.51 (m, 2 H), 1.52-1.60(m, 2 H), 1.68-1.98 (m, 10 H), (ES⁺) at methylcyclobutyl)amino] 2.08 (d,J = 8.5 Hz, 4 H), 2.13-2.32 (m, 2 H), 2.33-2.45 4.54 min, UV activepiperidin-1-yl}-6- (m, 2 H), 2.71-2.88 (m, 3 H), 2.89-3.03 (m, 2 H),3.26 azaspiro[3.4]octane-6- (s, 2 H), 3.35-3.43 (m, 3 H), 4.09 (q, J =6.7 Hz, 2 H) carboxylate 2-23 Isomer 2: Ethyl 2-(4- 4, 57 c (400 MHz,MeOD-d₄) δ: 1.23-1.36 (m, 6 H), 1.71-1.81 I m/z 408 (M + H)⁺{acetyl[(3-methyloxetan- and (m, 4 H), 1.87-2.03 (m, 6 H), 2.11-2.21 (m,2 H), 2.18 (ES⁺) at 3- 58 (s, 3H), 2.77-2.88 (m, 1 H), 2.95-3.07 (m, 3H), 3.27- 3.58 min, UV active yl)methyl]amino}piperidin- 3.31 (m 2 H),3.37-3.47 (m, 3 H), 3.68-3.79 (m, 1 H), 1-yl)-6- 4.13 (q, J = 7.1 Hz, 2H), 4.19 (d, J = 6.4 Hz, 2 H), 4.68- azaspiro[3.4]octane-6- 4.75 (m, 2H) carboxylate 2-24 Isomer 2: Ethyl 2-{4- 4 and 60 a (400 MHz, DMSO-d₆)δ: 1.16 (t, J = 7.0 Hz, 3 H), 1.42- I m/z 388 (M + H)⁺ [acetyl(2,2- 1.54(m, 1 H), 1.58-1.66 (m, 3 H), 1.66-1.92 (m, 6 H), (ES⁺) atdifluoroethyl)amino]piperi 1.92-2.05 (m, 3 H), 2.11 (s, 2 H), 2.63-2.70(m, 1 H), 3.99 min, UV active din-1-yl}-6- 2.78-2.91 (m, 2 H), 3.08-3.20(m, 2 H), 3.22-3.31 (m, 2 azaspiro[3.4]octane-6- H), 3.45-3.66 (m, 2 H),3.67-3.84 (m, 1 H), 4.00 (q, J = carboxylate 7.0 Hz, 2 H), 5.86-6.35 (m,1 H) 2-25 Isomer 2: Ethyl 2-{4- 4 and 62 a (400 MHz, DMSO-d₆) δ: 1.16(t, J = 7.0 Hz, 3 H), 1.51 (d, I m/z 406 (M + H)⁺ [acetyl(2,2,2- J =10.4 Hz, 1 H), 1.57-1.87 (m, 9 H), 1.96-2.02 (m, 2 (ES⁺) attrifluoroethyl)amino]piperi H), 2.05 (s, 1 H), 2.14 (s, 2 H), 2.56-2.72(m, 1 H), 2.75- 4.05 min, UV active din-1-yl}-6- 2.89 (m, 2 H), 3.14 (d,J = 5.5 Hz, 2 H), 3.26-3.32 (m, 2 azaspiro[3.4]octane-6- H), 3.57-3.87(m, 1 H), 3.94-4.12 (m, 3 H), 4.21 (q, J = carboxylate 9.0 Hz, 1 H) 2-26Isomer 1: Ethyl 2-{4- 4 and 64 a (400 MHz, MeOD-d₄) δ: 1.20-1.33 (m, 6H), 1.35 (s, 2 H), I m/z 398 (M + H)⁺ [acetyl(2-fluoro-2- 1.38 (s, 1 H),1.43 (s, 1 H), 1.67-1.82 (m, 2 H), 1.82- (ES⁺) atmethylpropyl)amino]piper 2.05 (m, 7 H), 2.06-2.19 (m, 3 H), 2.23 (s, 2H), 2.33- 3.94 min, UV active idin-1-yl}-6- 2.51 (m, 1 H), 2.78 (sxt, J= 8.3 Hz, 1 H), 2.92-3.06 (m, 2 azaspiro[3.4]octane-6- H), 3.37 (s, 3H), 3.42-3.80 (m, 3 H), 4.12 (q, J = 6.8 Hz, carboxylate 2 H) 2-26Isomer 2: Ethyl 2-{4- 4 and 64 a (400 MHz, MeOD-d₄) δ: 1.21-1.32 (m, 6H), 1.35 (s, 2 H), I m/z 398 (M + H)⁺ [acetyl(2-fluoro-2- 1.38 (s, 1 H),1.44 (s, 1 H), 1.70-1.86 (m, 2 H), 1.86- (ES⁺) atmethylpropyl)amino]piper 2.06 (m, 7 H), 2.06-2.19 (m, 3 H), 2.23 (s, 2H), 2.36- 4.05 min, UV active idin-1-yl}-6- 2.55 (m, 1 H), 2.73-2.94 (m,1 H), 3.02 (d, J = 6.4 Hz, 2 azaspiro[3.4]octane-6- H), 3.36-3.48 (m, 3H), 3.48-3.82 (m, 3 H), 4.11 (q, J = carboxylate 6.8 Hz, 2 H) 2-27Isomer 2: Ethyl 2-{4- 4 and 66 a (400 MHz, MeOD-d₄) δ: 1.24 (t, J = 7.0Hz, 3 H), 1.55- I m/z 402 (M + H)⁺ [acetyl(2,2- 1.64 (m, 2 H), 1.69-1.97(m, 10 H), 2.05-2.25 (m, 6 H), (ES⁺) at difluoropropyl)amino]pipe2.71-2.85 (m, 1 H), 2.97 (t, J = 10.1 Hz, 2 H), 3.26 (s, 2 3.95 min, UVactive ridin-1-yl}-6- H), 3.34-3.42 (m, 2 H), 3.68-3.86 (m, 3 H), 4.09(q, J = azaspiro[3.4]octane-6- 6.9 Hz, 2 H) carboxylate 2-28 Isomer 2:Ethyl 2-{4- 4 and 68 a (400 MHz, MeOD-d₄) δ: 1.25 (t, J = 7.0 Hz, 3 H),1.49- I m/z 414 (M + H)⁺ [acetyl(3,3- 2.01 (m, 9 H), 2.07-2.22 (m, 5 H),2.45-2.71 (m, 2 H), (ES⁺) at difluorocyclobutyl)amino] 2.80 (quin, J =7.9 Hz, 1 H), 2.97 (d, J = 11.0 Hz, 2 H), 3.85 min, UV activepiperidin-1-yl}-6- 3.22-3.27 (m, 2 H), 3.40 (q, J = 6.7 Hz, 3 H),3.48-3.89 azaspiro[3.4]octane-6- (m, 4 H), 4.10 (q, J = 7.1 Hz, 2 H)carboxylate 2-29 Isomer 1: Ethyl 2-{4- 4 and 70 a (400 MHz, MeOD-d₄) δ:1.15 (s, 3 H), 1.22 (s, 3 H), 1.24- I m/z 410 (M + H)⁺[acetyl(2-methoxy-2- 1.32 (m, 3 H), 1.66-1.80 (m, 3 H), 1.85-2.04 (m, 5H), (ES⁺) at methylpropyl)amino]piper 2.04-2.18 (m, 5 H), 2.21 (s, 1 H),2.50-2.68 (m, 1 H), 3.98 min, UV active iclin-1-yl}-6- 2.69-2.85 (m, 1H), 2.90-3.06 (m, 2 H), 3.23 (d, J = 2.9 azaspiro[3.4]octane-6- Hz, 3H), 3.35-3.44 (m, 5 H), 3.48-3.53 (m, 1 H), 3.58- carboxylate 3.72 (m, 1H), 4.12 (q, J = 6.8 Hz, 2 H) 2-29 Isomer 2: Ethyl 2-{4- 4 and 70 a (400MHz, MeOD-d₄) δ: 1.15 (s, 3 H), 1.22 (s, 3 H), 1.27 I m/z 410 (M + H)⁺[acetyl(2-methoxy-2- (t, J = 7.1 Hz, 3 H), 1.67-1.86 (m, 3 H), 1.86-1.99(m, 5 (ES⁺) at methylpropyl)amino]piper H), 2.03-2.19 (m, 5 H), 2.21 (s,1 H), 2.50-2.66 (m, 1 H), 4.12 min, UV active idin-1-yl}-6- 2.71-2.89(m, 1 H), 2.92-3.06 (m, 2 H), 3.23 (d, J = 3.9 azaspiro[3.4]octane-6-Hz, 3 H), 3.28 (br. s., 2 H), 3.38-3.42 (m, 4 H), 3.56- carboxylate 3.76(m, 1 H), 4.11 (q, J = 6.8 Hz, 2 H) 2-30 Isomer 1: Ethyl 2-(4- (400 MHz,DMSO-d₆) δ: 1.17 (t, J = 6.6 Hz, 3 H), 1.40- I m/z 422 (M + H)⁺{acetyl[(1- 1.69 (m, 4 H), 1.69-1.88 (m, 8 H), 1.90-2.05 (m, 4 H), (ES⁺)at methoxycyclobutyl)methyl] 4 and 71 a 2.10 (br. s., 2 H), 2.12-2.23(m, 1 H), 2.23-2.42 (m, 1 4.03 min, UV active amino}piperidin-1-yl)-6-H), 2.57-2.72 (m, 1 H), 2.73-2.95 (m, 2 H), 3.08-3.18azaspiro[3.4]octane-6- (m, 4 H), 3.18-3.29 (m, 4 H), 3.40-3.64 (m, 3 H),4.01 carboxylate (q, J = 7.3 Hz, 2 H) 2-30 Isomer 2: Ethyl 2-(4- 4 and71 a (400 MHz, DMSO-d₆) δ: 1.16 (t, J = 7.1 Hz, 3 H), 1.47- I m/z 422(M + H)⁺ {acetyl[(1- 1.56 (m, 4 H), 1.66-1.88 (m, 9 H), 1.90-2.20 (m, 7H), (ES⁺) at methoxycyclobutyl)methyl] 2.22-2.37 (m, 1 H), 2.55-2.70 (m,1 H), 2.76-2.83 (m, 2 4.16 min, UV active amino}piperidin-1-yl)-6- H),3.09-3.20 (m, 5 H), 3.27 (q, J = 6.5 Hz, 2 H), 3.40-azaspiro[3.4]octane-6- 3.60 (m, 3 H), 4.00 (d, J = 7.3 Hz, 2 H)carboxylate 2-31 Isomer 1: Ethyl 2-(4- 4 and 74 a (400 MHz, MeOD-d₄) δ:1.22-1.36 (m, 3 H), 1.56-2.28 I m/z 408 (M + H)⁺ {acetyl[(1- (m, 20 H),2.46-2.63 (m, 1 H), 2.74-2.91 (m, 1 H), 2.93- (ES⁺) athydroxycyclobutyl)methyl] 3.08 (m, 2 H), 3.36-3.44 (m, 4 H), 3.46-3.59(m, 2 H), 3.58 min, UV active amino}piperidin-1-yl)-6- 3.70-3.82 (m, 1H), 4.09-4.18 (m, 2 H). One azaspiro[3.4]octane-6- exchangeable protonnot observed. carboxylate 2-31 Isomer 2: Ethyl 2-(4- 4 and 74 a (400MHz, MeOD-d₄) δ: 1.28 (t, J = 7.0 Hz, 3 H), 1.57- I m/z 408 (M + H)⁺{acetyl[(1- 2.28 (m, 20 H), 2.45-2.59 (m, 1 H), 2.75-2.91 (m, 1 H),(ES⁺) at hydroxycyclobutyl)methyl] 2.92-3.08 (m, 2 H), 3.25-3.32 (m, 2H), 3.38-3.46 (m, 2 3.64 min, UV active amino}piperidin-1-yl)-6- H),3.48-3.61 (m, 2 H), 3.70-3.84 (m, 1 H), 4.13 (q, J =azaspiro[3.4]octane-6- 6.9 Hz, 2 H). One exchangeable proton notobserved. carboxylate 2-32 Isomer 2: Ethyl 2-{4- 4 and 76 a (400 MHz,MeOD-d₄) δ: 1.27 (t, J = 6.8 Hz, 3 H), 1.80 (d, I m/z 354 (M + H)⁺[acetyl(methoxy)amino]pi J = 11.7 Hz, 2 H), 1.84-2.03 (m, 8 H), 2.11 (d,J = 2.4 Hz, 2 (ES⁺) at peridin-1-yl}-6- H), 2.16 (s, 3 H), 2.80 (quin, J= 7.9 Hz, 1 H), 3.00 (d, J = 3.59 min, UV active azaspiro[3.4]octane-6-10.3 Hz, 2 H), 3.29 (s, 2 H), 3.41 (q, J = 6.5 Hz, 2 H), 3.82carboxylate (s, 3 H), 4.11 (q, J = 7.3 Hz, 2 H), 4.15-4.27 (m, 1 H) 2-33Isomer mixture: Ethyl 2- 4 and 78 a (400 MHz, DMSO-d₆) δ: 0.96-1.32 (m,5 H), 1.52-2.06 I m/z 400 (M + H)⁺ {4- (m, 10 H), 2.40-2.85 (m, 5 H),2.98-3.29 (m, 5 H), 3.83- (ES⁺) at [acetyl(phenyl)amino]pipe 4.16 (m, 2H), 4.25-4.61 (m, 1 H), 7.06-7.32 (m, 2 H), 4.09 min, UV activeridin-1-yl}-6- 7.32-7.61 (m, 3 H) azaspiro[3.4]octane-6- carboxylate2-34 Isomer mixture: Ethyl 2- 4 and 80 a (400 MHz, MeOD-d₄) δ: 1.20-1.38(m, 4 H), 1.38-1.56 I m/z 401 (M + H)⁺ {4-[acetyl(pyridin-2- (m, 2 H),1.72-2.01 (m, 12 H), 2.02-2.15 (m, 2 H), 2.69- (ES⁺) atyl)amino]piperidin-1-yl}-6- 2.81 (m, 1 H), 2.85-2.99 (m, 2 H), 3.21-3.45(m, 2 H), 3.39 min, UV active azaspiro[3.4]octane-6- 4.05-16 (m, 2 H),4.44-4.54 (m, 1 H), 7.42 (d, J = 7.9 carboxylate Hz, 1 H), 7.53 (ddd, J= 7.6, 5.0, 0.9 Hz, 1 H), 8.02 (td, J = 7.7, 2.0 Hz, 1 H), 8.61 (dt, J =4.4, 1.3 Hz, 1 H) 2-35 Isomer 2: Ethyl 2-{4- 4 and 82 a (400 MHz,DMSO-d₆) δ: 1.09-1.24 (m, 3 H), 1.38-1.60 I m/z 414 (M + H)⁺[acetyl(benzyl)amino]pipe (m, 4 H), 1.61-1.75 (m, 4 H), 1.75-1.87 (m, 2H), 1.91- (ES⁺) at ridin-1-yl}-6- 1.97 (m, 4 H), 2.18 (s, 2 H),2.71-2.79 (m, 3 H), 3.11 (dd, 4.14 min, UV active azaspiro[3.4]octane-6-J = 6.1, 3.4 Hz, 2 H), 3.20-3.31 (m, 2 H), 3.93-4.05 (m, carboxylate 2H), 4.43-4.63 (m, 2 H), 7.13-7.31 (m, 4 H), 7.32-7.42 (m, 1 H) 2-36Isomer 2: Ethyl 2-{4- 4 and 90 a (400 MHz, DMSO-d₆) δ: 1.16 (t, J = 7.1Hz, 3 H), 1.51- I m/z 405 (M + H)⁺ [acetyl(1,3-oxazol-5- 1.88 (m, 8 H),1.89-2.09 (m, 4 H), 2.40-2.69 (m, 5 H), (ES⁺) atylmethyl)amino]piperidin- 2.75-2.88 (m, 2 H), 3.07-3.19 (m, 2 H),3.20-3.29 (m, 2 3.38 min, UV active 1-yl}-6- H), 3.99 (q, J = 7.1 Hz, 2H), 4.39-4.66 (m, 2 H), 6.88- azaspiro[3.4]octane-6- 7.17 (m, 1 H),8.16-8.36 (m, 1 H) carboxylate 2-37 Isomer 2: Ethyl 2-{4- 4 and 93 a(400 MHz, DMSO-d₆) δ: 1.27 (t, J = 7.1 Hz, 3 H), 1.66- I m/z 405 (M +H)⁺ [acetyl(1,3-oxazol-2- 2.03 (m, 10 H), 2.04-2.29 (m, 5 H), 2.71-2.89(m, 1 H), (ES⁺) at ylmethyl)amino]piperidin- 2.96-3.05 (m, 2 H),3.21-3.45 (m, 4 H), 3.82-3.96 (m, 1 4.36 min, UV active 1-yl}-6- H),4.10 (q, J = 7.1 Hz, 2 H), 4.61-4.77 (m, 2 H), 7.03-azaspiro[3.4]octane-6- 7.25 (m, 1 H), 7.82-7.98 (m, 1 H) carboxylate2-38 Isomer 2: (1,1-2H2)-Ethyl 114, 3, d (400 MHz, MeOD-d₄) δ: 1.11-1.33(m, 6 H), 1.64-2.02 E m/z 354 (M + H)⁺ 2-{4- 115 and (m, 10 H),2.08-2.20 (m, 4 H), 2.75-2.86 (m, 1 H), 2.96- (ES⁺) at[acetyl(ethyl)amino]piperi 116 3.06 (m, 2 H), 3.29 (s, 2 H), 3.35-3.45(m, 4 H), 3.66- 3.37 min, UV active din-1-yl}-6- 3.77 (m, 1 H),4.21-4.34 (m, 1 H) azaspiro[3.4]octane-6- carboxylate 2-39 Isomer 2:Ethyl 2-{4- 4 and 94 a (400 MHz, MeOD-d₄) δ: 1.15(t, J = 7.0 Hz, 3 H),1.27(t, I m/z 368 (M + H)⁺ [ethyl(methoxycarbonyl) J = 7.1 Hz, 3 H),1.64-2.00 (m, 10 H), 2.13 (ddd, J = 9.7, (ES⁺) atamino]piperidin-1-yl}-6- 7.3, 2.6 Hz, 2 H), 2.80 (t, J = 8.0 Hz, 1 H),2.99 (d, J = 4.26 min, UV active azaspiro[3.4]octane-6- 11.5 Hz, 2 H),3.19-3.45 (m, 6 H), 3.70 (s, 3 H), 3.79- carboxylate 3.93 (m, 1 H), 4.11(q, J = 7.2 Hz, 2 H) 2-40 Isomer 2: Ethyl 2-{4- 4 and 95 a (400 MHz,DMSO-d₆) δ: 1.03 (t, J = 6.9 Hz, 3 H), 1.16 (t, I m/z 382 (M + H)⁺[(ethoxycarbonyl)(ethyl)a J = 7.2 Hz, 6 H), 1.46-1.81 (m, 6 H),1.94-2.04 (m, 4 H), (ES⁺) at mino]piperidin-1-yl}-6- 2.77-2.86 (m, 2 H),3.07-3.15 (m, 4 H), 3.16-3.49 (m, 6 4.52 min, UV activeazaspiro[3.4]octane-6- H), 4.01 (dd, J = 8.6, 6.9 Hz, 4 H) carboxylate2-41 Isomer 2: Ethyl 2-{4- 4 and 96 a (400 MHz, MeOD-d₄) δ: 1.18-1.31(m, 9 H), 1.52-1.63 I m/z 382 (M + H)⁺ [(methoxycarbonyl)(propan- (m, 2H), 1.82-2.04 (m, 6 H), 2.04-2.17 (m, 4 H), 2.80 (t, (ES⁺) at2-yl)amino]piperidin-1- J = 7.5 Hz, 1 H), 2.89-3.00 (m, 2 H), 3.24-3.44(m, 6 H), 4.36 min, UV active yl}-6-azaspiro[3.4]octane- 3.69 (s, 3 H),4.11 (q, J = 7.1 Hz, 2 H) 6-carboxylate 2-42 Isomer 2: Ethyl 2-{4- 4 and98 a (400 MHz, MeOD-d₄) δ: 1.22-1.30 (m, 3 H), 1.72-1.80 I m/z 422 (M +H)⁺ [(methoxycarbonyl)(2,2,2- (m, 2 H), 1.82-2.01 (m, 8 H), 2.07-2.17(m, 2 H), 2.73- (ES⁺) at trifluoroethyl)amino]piperi 2.85 (m, 1 H),2.94-3.04 (m, 2 H), 3.24-3.29 (m, 2 H), 4.67 min, UV active din-1-yl}-6-3.36-3.45 (m, 2 H), 3.58-3.70 (m, 1 H), 3.70-3.80 (m, 3azaspiro[3.4]octane-6- H), 3.95-4.06 (m, 2 H), 4.07-4.16 (m, 2 H)carboxylate 2-43 Isomer 2: Ethyl 2-(4-{[(2- 4 and 99 a (400 MHz,MeOD-d₄) δ: 1.27 (t, J = 7.1 Hz, 3 H), 1.46- I m/z 467 (M + H)⁺methoxyethoxy)carbonyl] 1.60 (m, 2 H), 1.76-2.00 (m, 9 H), 2.08-2.18 (m,2 H), (ES⁺) at (2,2,2- 2.59-2.89 (m, 4 H), 2.93-3.02 (m, 2 H), 3.17-3.50(m, 4.56 min, UV active trifluoroethyl)amino}piperi 10 H), 4.11 (d, J =7.1 Hz, 2 H) din-1-yl)-6- azaspiro[3.4]octane-6- carboxylate 2-44 Isomer2: Ethyl 2-{4- 4 and a (400 MHz, MeOD-d₄) δ: 1.10 (t, J = 6.8 Hz, 3 H),1.27 (t, I m/z 381 (M + H)⁺ [(dimethylcarbamoyl)(ethyl) 101 J = 7.1 Hz,3 H), 1.79-1.91 (m, 2 H), 1.92-2.18 (m, 6 H), (ES⁺) atamino]piperidin-1-yl}-6- 2.21-2.32 (m, 2 H), 2.35-2.52 (m, 2 H), 2.87(s, 6 H), 3.92 min, UV active azaspiro[3.4]octane-6- 3.12-3.57 (m, 10H), 4.12 (d, J = 7.1 Hz, 2 H) carboxylate 2-45 Isomer 2: Ethyl 2-{4- 4and a (400 MHz, DMSO-d₆) δ: 0.96 (t, J = 7.1 Hz, 3 H), 1.16 (t, I m/z356 (M + H)⁺ [ethyl(2- 103 J = 7.1 Hz, 3 H), 1.27-1.45 (m, 2 H),1.49-2.06 (m, 10 H), (ES⁺) at fluoroethyl)amino]piperidin- 2.24-2.90 (m,8 H), 3.04-3.34 (m, 6 H), 3.99 (d, J = 7.1 4.17 min, UV active 1-yl}-6-Hz, 2 H) azaspiro[3.4]octane-6- carboxylate 2-46 Isomer 2: Ethyl 2-{4- 4and a (400 MHz, DMSO-d₆) δ: 0.95 (t, J = 7.1 Hz, 3 H), 1.10- I m/z 375(M + H)⁺ [(2,2- 105 1.22 (m, 3 H), 1.26-1.41 (m, 2 H), 1.57-1.80 (m, 8H), (ES⁺) at difluoroethyl)(ethyl)amino] 1.93-2.06 (m, 2 H), 2.37-2.66(m, 5 H), 2.69-2.85 (m, 4 4.56 min, UV active pipendin-1-yl}-6- H),3.15-3.30 (m, 4 H), 4.01 (q, J = 7.1 Hz, 2 H) azaspiro[3.4]octane-6-carboxylate 2-47 Isomer 2: Ethyl 2-{4- 4, 57, c (400 MHz, MeOD-d₄) δ:1.28 (t, J = 7.3 Hz, 3 H), 1.62- I m/z 396 (M + H)⁺[(methoxycarbonyl)(oxetan- 106 1.72 (m, 2 H), 1.89-2.06 (m, 6 H),2.09-2.20 (m, 2 H), (ES⁺) at 3-yl)amino]piperidin-1- and 40 2.76-2.90(m, 1 H), 2.96-3.04 (m, 2 H), 3.30 (s, 2 H), 3.96 min, UV activeyl}-6-azaspiro[3.4]octane- 3.37-3.48 (m, 4 H), 3.62-3.73 (m, 1 H), 3.75(s, 3 H), 6-carboxylate 4.13 (q, J = 7.1 Hz, 2 H), 4.71-4.78 (m, 2 H),4.81-5.00 (m, 3 H) 2-48 Isomer 2: Ethyl 2-{4- 4 and a (400 MHz, DMSO-d₆)δ: 0.33-0.52 (m, 4 H), 1.16 (t, J = I m/z 404 (M + H)⁺[cyclopropyl(2,2,2- 109 7.1 Hz, 3 H), 1.42-1.88 (m, 10 H), 1.92-2.02 (m,2 H), (ES⁺) at trifluoroethyl)amino]piperi 2.13-2.23 (m, 1 H), 2.54-2.67(m, 2 H), 2.76-2.86 (m, 2 5.47 min, UV active din-1-yl}-6- H), 3.14 (d,J = 5.8 Hz, 2 H), 3.22-3.36 (m, 4 H), 3.99 (q, azaspiro[3.4]octane-6- J= 7.1 Hz, 2 H) carboxylate 2-49 Isomer 2: Ethyl 2-{4- 4 and a (400 MHz,MeOD-d₄) δ: 0.40-0.63 (m, 4 H), 0.87-0.99 I m/z 366 (M + H)⁺[cyclopropyl(2- 111 (m, 1 H), 1.19-1.36 (m, 3 H), 1.57-1.78 (m, 3 H),1.82- (ES⁺) at hydroxyethyl)amino]piperi 2.05 (m, 8 H), 2.07-2.25 (m, 2H), 2.77-2.88 (m, 4 H), 3.76 min, UV active din-1-yl}-6- 2.94-3.07 (m, 2H), 3.25-3.55 (m, 4 H), 3.60-3.75 (m, 2 azaspiro[3.4]octane-6- H), 4.11(d, J = 7.1 Hz, 2 H) carboxylate 2-50 Isomer 2: Ethyl 2-{4- 4 and a (400MHz, DMSO-d₆) δ: 1.16 (t, J = 7.1 Hz, 3 H), 1.21- I m/z 380 (M + H)⁺[cyclobutyl(2- 113 1.43 (m, 3 H), 1.46-1.67 (m, 5 H), 1.67-2.02 (m, 10H), (ES⁺) at hydroxyethyl)amino]piperi 2.30-2.49 (m, 4 H), 2.65-2.71 (m,1 H), 2.75-2.83 (m, 2 3.95 min, UV active din-1-yl}-6- H), 3.11-3.17 (m,2 H), 3.23-3.33 (m, 4 H), 3.99 (d, J = azaspiro[3.4]octane-6- 7.1 Hz, 2H), 4.26-4.32 (m, 1 H) carboxylate 2-51 Isomer 2: Ethyl 2-{4-[(2- 4 anda (400 MHz, MeOD-d₄) δ: 1.28 (t, J = 7.1 Hz, 3 H), 1.54- I m/z 408 (M +H)⁺ hydroxyethyl)(2,2,2- 117 1.69 (m, 2 H), 1.83-1.94 (m, 2 H),1.93-2.16 (m, 6 H), (ES⁺) at trifluoroethyl)amino]piperi 2.15-2.25 (m, 2H), 2.69-2.79 (m, 1 H), 2.82 (t, J = 6.6 4.25 min, UV activedin-1-yl}-6- Hz, 2 H), 2.95-3.06 (m, 1 H), 3.06-3.18 (m, 2 H), 3.20-azaspiro[3.4]octane-6- 3.33 (m, 4 H), 3.44 (q, J = 7.1 Hz, 2 H), 3.60(t, J = 6.6 Hz, carboxylate 2 H), 4.14 (q, J = 7.2 Hz, 2 H). Oneexchangeable proton not observed. 2-52 Isomer 2: Ethyl 2-{4-[(2- 4 and a(400 MHz, MeOD-d₄) δ: 1.28 (t, J = 7.1 Hz, 3 H), 1.48- I m/z 422 (M +H)⁺ methoxyethyl)(2,2,2- 119 1.60 (m, 2 H), 1.78-1.88 (m, 4 H),1.88-2.02 (m, 4 H), (ES⁺) at trifluoroethyl)amino]piperi 2.09-2.18 (m, 2H), 2.61-2.72 (m, 1 H), 2.73-2.83 (m, 1 4.97 min, UV active din-1-yl}-6-H), 2.86 (t, J = 6.2 Hz, 2 H), 2.98 (d, J = 12.0 Hz, 2 H),azaspiro[3.4]octane-6- 3.19-3.31 (m, 4 H), 3.35 (s, 3H), 3.42 (q, J =7.0 Hz, 2 carboxylate H), 3.47 (t, J = 6.1 Hz, 2 H), 4.12 (q, J = 7.1Hz, 2 H) 2-53 Isomer 2: Ethyl 2-{4- 4 and a (400 MHz, MeOD-d₄) δ: 1.07(t, J = 7.1 Hz, 3 H), 1.28 (t, I m/z 366 (M + H)⁺ [ethyl(oxetan-3- 120 J= 7.1 Hz, 3 H), 1.39-1.59 (m, 2 H), 1.65-1.78 (m, 2 H), (ES⁺) atyl)amino]piperidin-1-yl}-6- 1.81-2.02 (m, 6 H), 2.09-2.18 (m, 2 H),2.51-2.64 (m, 1 3.66 min, UV active azaspiro[3.4]octane-6- H), 2.73 (q,J = 7.0 Hz, 2 H), 2.76-2.86 (m, 1 H), 2.99 (d, carboxylate J = 11.7 Hz,2 H), 3.29 (s, 2 H), 3.41 (q, J = 6.8 Hz, 2 H), 4.09-4.23 (m, 3 H),4.64-4.71 (m, 4 H) 2-54 Isomer 2: Ethyl 2-{4- 4 and a (400 MHz, MeOD-d₄)δ: 1.28 (t, J = 7.1 Hz, 3 H), 1.39- I m/z 420 (M + H)⁺[oxetan-3-yl(2,2,2- 122 1.52 (m, 2 H), 1.72 (d, J = 12.8 Hz, 2 H),1.78-2.00 (m, 6 (ES⁺) at trifluoroethyl)amino]piperi H), 2.09-2.18 (m, 2H), 2.60-2.71 (m, 1 H), 2.72-2.84 4.49 min, UV active din-1-yl}-6- (m, 1H), 2.97 (d, J = 9.8 Hz, 2 H), 3.29 (s, 2 H), 3.41 (q,azaspiro[3.4]octane-6- J = 6.4 Hz, 2 H), 3.50 (q, J = 9.5 Hz, 2 H), 4.12(q, J = 6.8 carboxylate Hz, 2 H), 4.33-4.40 (m, 1 H), 4.63 (t, J = 6.6Hz, 2 H), 4.74 (t, J = 7.2 Hz, 2 H) 2-55 Isomer 2: Ethyl 2-{4-[(2- 4 anda (400 MHz, MeOD-d₄) δ: 1.09 (t, J = 7.1 Hz, 3 H), 1.28 (t, I m/z 363(M + H)⁺ cyanoethyl)(ethyl)amino] 124 J = 7.1 Hz, 3 H), 1.50-1.63 (m, 2H), 1.79-1.89 (m, 4 H), (ES⁺) at piperidin-1-yl}-6- 1.89-2.02 (m, 4 H),2.10-2.20 (m, 2 H), 2.55 (t, J = 6.8 4.20 min, UV activeazaspiro[3.4]octane-6- Hz, 2 H), 2.65 (q, J = 7.2 Hz, 2 H), 2.73-2.86(m, 4 H), carboxylate 2.98 (d, J = 12.0 Hz, 2 H), 3.29 (s, 2 H), 3.42(q, J = 6.9 Hz, 2 H), 4.12 (q, J = 7.1 Hz, 2 H) 2-56 Isomer 2: Ethyl2-(4- 4 and a (400 MHz, MeOD-d₄) δ: 1.10 (t, J = 7.2 Hz, 3 H), 1.28 (t,I m/z 416 (M + H)⁺ {ethyl[2- 126 J = 7.0 Hz, 3 H), 1.54-1.67 (m, 2 H),1.77-2.02 (m, 8 H), (ES⁺) at (methylsulfonyl)ethyl]ami 2.09-2.19 (m, 2H), 2.58-2.70 (m, 3 H), 2.74-2.84 (m, 1 3.74 min, UV activenolpiperidin-1-yl)-6- H), 2.95-3.08 (m, 7 H), 3.22-3.31 (m, 4 H), 3.42(q, J = azaspiro[3.4]octane-6- 6.5 Hz, 2 H), 4.12 (q, J = 7.0 Hz, 2 H)carboxylate 2-57 Isomer 2: Ethyl 2-{4- 4 and a (400 MHz, MeOD-d₄) δ:1.17(t, J = 7.0 Hz, 3 H), 1.28(t, I m/z 340 (M + H)⁺[ethyl(methoxy)amino]pip 128 J = 7.1 Hz, 3 H), 1.57-1.69 (m, 2 H),1.82-2.00 (m, 8 H), (ES⁺) at eridin-1-yl}-6- 2.09-2.17 (m, 2 H),2.56-2.66 (m, 1 H), 2.74-2.86 (m, 3 4.30 min, UV activeazaspiro[3.4]octane-6- H), 2.96 (d, J = 11.7 Hz, 2 H), 3.29 (s, 2 H),3.42 (q, J = carboxylate 6.8 Hz, 2 H), 3.56 (s, 3H), 4.12 (q, J = 6.8Hz, 2 H) 2-58 Isomer 2: Ethyl 2-{4-[(2- 4 and a (400 MHz, DMSO-d₆) δ:1.18 (t, J = 7.1 Hz, 3 H), 1.56- I m/z 402 (M + H)⁺hydroxyethyl)(phenyl)ami 130 1.70 (m, 4 H), 1.74-1.90 (m, 6 H),1.91-2.08 (m, 2 H), (ES⁺) at no]piperidin-1-yl}-6- 2.64-2.73 (m, 1 H),2.86 (d, J = 11.7 Hz, 2 H), 3.17 (d, 4.72 min, UV activeazaspiro[3.4]octane-6- J = 4.1 Hz, 2 H), 3.19-3.35 (m, 4 H), 3.40-3.48(m, 2H), carboxylate 3.49-3.59 (m, 1 H), 4.01 (q, J = 7.1 Hz, 2 H), 4.78(br. s, 1 H), 6.59 (t, J = 7.2 Hz, 1 H), 6.75 (d, J = 8.3 Hz, 2 H),7.12-7.18 (m, 2 H) 2-59 Isomer 2: Ethyl 2-{4- 4 and a (400 MHz, DMSO-d₆)δ: 1.17 (t, J = 7.0 Hz, 3 H), 1.36- I m/z 454 (M + H)⁺ [benzyl(2,2,2-132 1.50 (m, 2 H), 1.49-1.61 (m, 2 H), 1.63-1.77 (m, 4 H), (ES⁺) attrifluoroethyl)amino]piperi 1.77-1.87 (m, 2 H), 1.91-2.00 (m, 2 H),2.32-2.44 (m, 1 5.89 min, UV active din-1-yl}-6- H), 2.56-2.62 (m, 1 H),2.81 (d, J = 11.3 Hz, 2 H), 3.13 azaspiro[3.4]octane-6- (d, J = 5.5 Hz,2 H), 3.23-3.33 (m, 4 H), 3.82 (s, 2 H), carboxylate 4.00 (q, J = 7.0Hz, 2 H), 7.21-7.28 (m, 1 H), 7.30-7.36 (m, 4 H) 2-60 Isomer 2: Ethyl2-{4- 4 and a (400 MHz, DMSO-d₆) δ: 0.96 (t, J = 7.0 Hz, 3 H), 1.18 (t,I m/z 391 (M + H)⁺ [ethyl(1,3-oxazol-5- 133 J = 7.2 Hz, 3 H), 1.35-1.47(m, 2 H), 1.60-1.72 (m, 4 H), (ES⁺) at ylmethyl)amino]piperidin-1.73-1.90 (m, 4 H), 1.94-2.03 (m, 2 H), 2.37-2.46 (m, 2 3.85 min, UVactive 1-yl}-6- H), 2.59-2.68 (m, 2 H), 2.77-2.86 (m, 2 H), 3.10-3.22azaspiro[3.4]octane-6- (m, 2 H), 3.25-3.33 (m, 2 H), 3.71 (s, 2 H), 4.01(q, J = carboxylate 7.0 Hz, 2 H), 7.02 (s, 1 H), 8.27 (s, 1 H) 2-61Isomer 2: Ethyl 2-{4-[(2- 4 and a (400 MHz, MeOD-d₄) δ: 1.27 (t, J = 7.2Hz, 3 H), 1.53- I m/z 409 (M + H)⁺ fluoroethyl)(1,3-oxazol-5- 135 1.68(m, 2 H), 1.81-2.03 (m, 8 H), 2.08-2.19 (m, 2 H), (ES⁺) atylmethyl)amino]piperidin- 2.57-2.68 (m, 1 H), 2.73-2.84 (m, 1 H), 2.89(t, J = 5.3 3.82 min, UV active 1-yl}-6- Hz, 1 H), 2.92-3.03 (m, 3 H),3.29 (s, 2 H), 3.41 (q, J = azaspiro[3.4]octane-6- 6.9 Hz, 2 H), 3.92(s, 2 H), 4.12 (q, J = 7.1 Hz, 2 H), 4.37 carboxylate (t, J = 5.3 Hz, 1H), 4.49 (t, J = 5.3 Hz, 1 H), 7.06 (s, 1 H), 8.18 (s, 1 H) 2-62 Isomer2: Ethyl 2-{4- 4 and a (400 MHz, MeOD-d₄) δ: 1.27 (t, J = 7.0 Hz, 3 H),1.52- I m/z 445 (M + H)⁺ [(1,3-oxazol-5- 136 1.66 (m, 2 H), 1.82-2.02(m, 8 H), 2.12-2.20 (m, 2 H), (ES⁺) at ylmethyl)(2,2,2- 2.65-2.77 (m, 1H), 2.81-2.93 (m, 1 H), 2.97-3.08 (m, 2 4.47 min, UV activetrifluoroethyl)amino]piperi H), 3.25-3.32 (m, 4 H), 3.41 (q, J = 6.9 Hz,2 H), 4.00 (s, din-1-yl}-6- 2 H), 4.12 (q, J = 7.0 Hz, 2 H), 7.07 (s, 1H), 8.19 (s, 1 H) azaspiro[3.4]octane-6- carboxylate 2-63 Isomer 2b:Ethyl 2-(4-{1- 4 and e (400 MHz, MeOD-d₄) δ: 0.82-0.97 (m, 3 H),1.12-1.42 I m/z 394 (M + H)⁺ [acetyl(ethyl)amino]propyl} 138 (m, 10 H),1.43-2.01 (m, 10 H), 2.03-2.25 (m, 5 H), 2.69- (ES⁺) atpiperidin-1-yl)-6- 2.83 (m, 1 H), 2.85-3.06 (m, 2 H), 3.14-3.33 (m, 4H), 3.99 min, UV active azaspiro[3.4]octane-6- 3.37-3.46 (m, 2 H), 4.12(q, J = 6.8 Hz, 2 H) carboxylate 2-64 Isomer 2: Ethyl 2-(4-{2- 4 and a(400 MHz, MeOD-d₄) δ: 1.20-1.31 (m, 6 H), 1.31-1.43 I m/z 394 (M + H)⁺[acetyl(ethyl)amino]propan- 140 (m, 2 H), 1.43 (s, 6 H), 1.50-1.57 (m, 2H), 1.71-1.80 (ES⁺) at 2-yl}piperidin-1-yl)-6- (m, 2 H), 1.88-2.00 (m, 4H), 2.08-2.20 (m, 5 H), 2.69- 4.01 min, UV active azaspiro[3.4]octane-6-2.82 (m, 2 H), 2.94-3.03 (m, 2 H), 3.29 (s, 2 H), 3.37- carboxylate 3.47(m, 4 H), 4.12 (q, J = 6.8 Hz, 2 H) 2-65 Isomer 1a: Ethyl 2-{4-[1- 4 andf (400 MHz, MeOD-d₄) δ: 1.08-1.42 (m, 7 H), 1.52 (d, J = I m/z 361 (M +H)⁺ (1H-pyrazol-1- 144 6.8 Hz, 3 H), 1.68-1.97 (m, 8 H), 2.05-2.21 (m, 2H), (ES⁺) at yl)ethyl]piperidin-1-yl}-6- 2.72-2.82 (m, 1 H), 2.82-2.91(m, 1 H), 2.95-3.03 (m, 1 3.87 min, UV active azaspiro[3.4]octane-6- H),3.35-3.41 (m, 3 H), 4.09-4.20 (m, 3 H), 6.30 (t, J = carboxylate 2.2 Hz,1 H), 7.52 (d, J = 2.0 Hz, 1 H), 7.67 (d, J = 2.0 Hz, 1 H) 2-65 Isomer2a: Ethyl 2-{4-[1- 4 and f (400 MHz, MeOD-d₄) δ: 1.06-1.43 (m, 7 H),1.52 (d, J = I m/z 361 (M + H)⁺ (1H-pyrazol-1- 144 6.4 Hz, 3 H),1.64-2.02 (m, 8 H), 2.04-2.19 (m, 2 H), (ES⁺) atyl)ethyl]piperidin-1-yl}-6- 2.69-2.81 (m, 1 H), 2.81-2.91 (m, 1 H),2.93-3.02 (m, 1 4.18 min, UV active azaspiro[3.4]octane-6- H), 3.37-3.46(m, 3 H), 4.06-4.22 (m, 3 H), 6.30 (t, J = carboxylate 2.0 Hz, 1 H),7.52 (d, J = 2.0 Hz, 1 H), 7.66 (d, J = 2.0 Hz, 1 H) 2-66 Isomer 2:Ethyl 2-[4-(1- 4 and a (400 MHz, DMSO-d₆) δ: 1.17 (t, J = 7.2 Hz, 3 H),1.31 (d, I m/z 387 (M + H)⁺ phenylethoxy)piperidin-1- 147 J = 6.4 Hz, 3H), 1.35-1.51 (m, 2 H), 1.58-1.70 (m, 1 H), (ES⁺) atyl]-6-azaspiro[3.4]octane- 1.71-1.92 (m, 6 H), 1.92-2.02 (m, 2 H),2.44-2.67 (m, 4 5.29 min, UV active 6-carboxylate H), 3.11-3.22 (m, 3H), 3.27 (q, J = 1.0 Hz, 2 H), 4.00 (q, J = 7.0 Hz, 2 H), 4.62 (q, J =6.3 Hz, 1 H), 7.23-7.30 (m, 1 H), 7.31-7.40 (m, 4 H) 2-67 Isomer 2:Ethyl 2-[4- 4 and a (400 MHz, DMSO-d₆) δ: 1.17 (t, J = 7.1 Hz, 3 H),1.35- I m/z 389 (M + H)⁺ (benzylsulfanyl)piperidin- 150 1.49 (m, 2 H),1.68-1.91 (m, 8 H), 1.92-2.02 (m, 2 H), (ES⁺) at 1-yl]-6- 2.56-2.71 (m,4 H), 3.15 (d, J = 6.6 Hz, 2 H), 3.28 (q, J = 5.37 min, UV activeazaspiro[3.4]octane-6- 6.9 Hz, 2 H), 3.78 (s, 2 H), 4.00 (q, J = 7.1 Hz,2 H), 7.21- carboxylate 7.28 (m, 1 H), 7.28-7.36 (m, 4 H) 3-1 Racemic:Ethyl 6-{4- 8 and 30 a (400 MHz, CDCl₃) δ: 0.80-0.94 (m, 4 H), 1.25 (t,J = 7.0 I m/z 364 (M + H)⁺ [acetyl(cyclopropyl)amino] Hz, 3 H),1.50-1.58 (m, 1 H), 1.69-2.20 (m, 12 H), 2.23 (ES⁺) atpiperidin-1-yl}-2- (s, 3 H), 2.46-2.62 (m, 2 H), 2.99-3.09 (m, 2 H),3.76- 3.60 min, UV active azaspiro[3.4]octane-2- 3.91 (m, 4 H), 4.11 (q,J = 7.0 Hz, 2 H) carboxylate 3-2 Isomer 2: Methyl 6-{4- 2 and g (400MHz, MeOD-d₄) δ: 1.09-1.29 (m, 3 H), 1.48-1.62 I m/z 338 (M + H)⁺[acetyl(ethyl)amino]piperi 152 (m, 1 H), 1.64-2.02 (m, 8 H), 2.03-2.26(m, 6 H), 2.61- (ES) at din-1-yl}-2- 2.74 (m, 1 H), 3.07-3.19 (m, 2 H),3.28-3.44 (m, 2 H), 3.11 min, UV active azaspiro[3.4]octane-2- 3.67 (s,3 H), 3.76-3.98 (m, 4 H), 4.21-4.33 (m, 1 H) carboxylate 3-3 Isomer 2:Ethyl 6-{4- 2 and g (400 MHz, MeOD-d₄) δ: 1.11-1.29 (m, 6 H), 1.49-1.62I m/z 352 (M + H)⁺ [acetyl(ethyl)amino]piperi 153 (m, 1 H), 1.65-2.03(m, 8 H), 2.04-2.13 (m, 1 H), 2.15 (ES⁺) at din-1-yl}-2- (d, J = 4.6 Hz,3 H), 2.16-2.24 (m, 2 H), 2.61-2.74 (m, 1 3.43 min, UV activeazaspiro[3.4]octane-2- H), 3.08-3.18 (m, 2 H), 3.29-3.42 (m, 2 H),3.76-3.96 carboxylate (m, 4 H), 4.10 (q, J = 7.2 Hz, 2 H), 4.22-4.32 (m,1 H)

Biological Activity Example A

Phospho-ERK1/2 Assays

Functional assays were performed using the Alphascreen Surefirephospho-ERK1/2 assay (Crouch & Osmond, Comb. Chem. High ThroughputScreen, 2008). ERK1/2 phosphorylation is a downstream consequence ofboth Gq/11 and Gi/o protein coupled receptor activation, making ithighly suitable for the assessment of M₁, M₃ (Gq/11 coupled) and M₂, M₄receptors (Gi/o coupled), rather than using different assay formats fordifferent receptor subtypes. CHO cells stably expressing the humanmuscarinic M₁, M₂, M₃ or M₄ receptor were plated (25K/well) onto 96-welltissue culture plates in MEM-alpha+10% dialysed FBS. Once adhered, cellswere serum-starved overnight. Agonist stimulation was performed by theaddition of 5 μL agonist to the cells for 5 min (37° C.). Media wasremoved and 50 μL of lysis buffer added. After 15 min, a 4 μL sample wastransferred to 384-well plate and 7 μL of detection mixture added.Plates were incubated for 2 h with gentle agitation in the dark and thenread on a PHERAstar plate reader. pEC₅₀ and E_(max) figures werecalculated from the resulting data for each receptor subtype.

The results are set out in Table 4 below.

For each example containing the 6-azaspiro[3.4]octane ring system twodiastereomers exist which have been separated, unless stated otherwise,and assigned (Isomer 1, Isomer 2) based on their analytical LCMSretention time. In most examples, Isomer 1 is not active. Where further(chiral) isomers exist, these have sometimes been separated and assigned(Isomer 1a, Isomer 1b) based on their chiral separation retention time.

For each example containing the 2-azaspiro[3.4]octane ring system twoenantiomers exist which have been separated, unless stated otherwise,and assigned (Isomer 1, Isomer 2) based on their chiral separationretention time.

Analytical data for active isomers is reported in Table 3. Data forseveral weakly active compounds are included in Table 4 to highlightpreference of absolute stereochemistry.

TABLE 4 Muscarinic Activity pEC₅₀ M1 pEC₅₀ M2 pEC₅₀ M3 pEC₅₀ M4 (% Emaxcf. (% Emax cf. (% Emax cf. (% Emax cf. Ex. No. ACh) ACh) ACh) ACh) ACh 8.3 (102)   7.8 (105)   8.1 (115)  8.1 (110) 1-1 5.6 (72) NT NT 6.5(77) 1-2 5.2 (59) NT NT  6.6 (103) 2-1 Isomer 2 6.3 (88) <4.7 (15) <4.7(13)  7.9 (108) 2-2 Isomer 2 6.2 (91) <4.7 (18) <4.7 (9)  7.4 (99) 2-3Isomer 1 5.7 (78) NT NT <4.7 (13)   2-3 Isomer 2 6.7 (93) <4.7 (13) <4.7(6)  7.5 (93) 2-4 Isomer 2  6.7 (104) <4.7 (3)  <4.7 (9)   7.5 (114) 2-5Isomer 2 6.2 (57) <4.7 (5)  <4.7 (5)  7.0 (91) 2-6 Isomer 2  7.0 (101)<4.7 (7)  <4.7 (14) 7.5 (98) 2-7 Isomer 2  7.1 (120) <4.7 (16) <4.7 (10) 7.9 (118) 2-8 Isomer 2 <4.7 (4)    <4.7 (6)  <4.7 (6)  7.7 (57) 2-9Isomer 2 <4.7 (14)   <4.7 (6)  <4.7 (5)  7.0 (92) 2-10 Isomer 2 6.1 (42)<4.7 (11) <4.7 (7)   7.5 (122) 2-11 Isomer 2 <4.7 (9)    NT NT 6.5 (25)2-12 Isomer 2 <4.7 (6)    NT NT 6.7 (41) 2-13 Isomer 2 <4.7 (18)   <4.7(5)  <4.7 (4)  6.9 (88) 2-14 Isomer 2 5.6 (47) <4.7 (12) <4.7 (5)   7.0(108) 2-15 Isomer 2  6.3 (117) NT NT 6.7 (95) 2-16 Isomer 2 6.4 (89)<4.7 (5)  <4.7 (2)  7.4 (91) 2-17 Isomer 1 <4.7 (27)   NT NT 5.9 (87)2-17 Isomer 2 5.9 (80) NT NT 6.7 (92) 2-18 Isomer 2 5.7 (89) NT NT  6.7(107) 2-19 Isomer 1 5.9 (32) NT NT 6.6 (62) 2-19 Isomer 2  7.0 (115)<4.7 (31) <4.7 (58)  7.2 (118) 2-20 Isomer 1 <4.7 (17)   NT NT 6.0 (36)2-20 Isomer 2  6.7 (113) <4.7 (60) <4.7 (33)  7.4 (108) 2-21 Isomer 14.8 (51) NT NT 6.5 (46) 2-21 isomer 2 7.2 (89) <4.7 (18) <4.7 (22) 6.8(93) 2-22 Isomer 2  6.5 (132) NT NT  7.1 (139) 2-23 Isomer 2 5.9 (43) NTNT 6.2 (65) 2-24 Isomer 2 6.4 (91) <4.7 (5)  <4.7 (17)  8.0 (124) 2-25Isomer 2  5.4 (125) NT NT  7.0 (101) 2-26 Isomer 1 <4.7 (23)   NT NT 6.0(52) 2-26 Isomer 2  6.4 (109) NT NT  7.2 (114) 2-27 Isomer 2  6.2 (108)NT NT  7.1 (105) 2-28 Isomer 2  6.8 (124) NT NT 6.6 (80) 2-29 Isomer 1<4.7 (17)   NT NT 5.9 (56) 2-29 Isomer 2 6.7 (99)   5.9 (29) <4.7 (73)6.8 (84) 2-30 Isomer 1 5.9 (33) NT NT 6.9 (59) 2-30 Isomer 2  7.0 (104)  5.3 (42) <4.7 (26) 7.1 (96) 2-31 Isomer 1 5.4 (27) NT NT 6.7 (62) 2-31Isomer 2 6.4 (78) NT NT 7.4 (95) 2-32 Isomer 2 5.6 (82) <4.7 (9)  <4.7(5)   7.4 (115) 2-33 Isomer 6.7 (97) NT NT 6.7 (78) mixture 2-34 Isomer 6.8 (116) NT NT 6.2 (56) mixture 2-35 Isomer 2 7.4 (85) <4.7 (21) <4.7(34) 6.9 (72) 2-36 Isomer 2 5.6 (45) NT NT 6.2 (71) 2-37 Isomer 2 5.9(26) NT NT 6.2 (55) 2-38 Isomer 2 6.2 (76) <4.7 (21) <5.2 (1)   7.7(115) 2-39 Isomer 2 6.5 (39) <4.7 (12) <4.7 (25)  7.6 (126) 2-40 Isomer2 6.3 (39) <4.7 (32) <4.7 (9)   7.2 (110) 2-41 Isomer 2  6.5 (109) <4.7(57) <4.7 (1)   7.8 (127) 2-42 Isomer 2  5.8 (120) <4.7 (21) <4.7 (39) 7.0 (112) 2-43 Isomer 2 5.6 (35) NT NT 6.9 (91) 2-44 isomer 2 5.6 (55)NT NT 6.4 (80) 2-45 Isomer 2 6.2 (30) <4.7 (9)  <4.7 (16) 7.2 (45) 2-46Isomer 2 6.5 (63) <4.7 (5)  <4.7 (12) 8.1 (98) 2-47 Isomer 2  6.4 (113)  6.3 (29)   5.9 (63) 7.2 (90) 2-48 Isomer 2  7.4 (117)   4.9 (68)   5.1(53) 7.4 (75) 2-49 Isomer 2 <4.7 (8)    NT NT 5.9 (78) 2-50 Isomer 2<4.7 (17)   NT NT 6.6 (70) 2-51 Isomer 2 6.5 (84) <4.7 (18) <4.7 (22)7.7 (89) 2-52 Isomer 2 6.4 (93) <4.7 (7)  <4.7 (7)   7.6 (106) 2-53Isomer 2 <4.7 (42)   NT NT 6.6 (62) 2-54 Isomer 2  6.6 (106) <4.7 (28)<4.7 (20) 7.5 (98) 2-55 Isomer 2 <4.7 (161) <4.7 (15) <4.7 (5)  7.6 (65)2-56 Isomer 2 <4.7 (7)    NT NT 5.9 (41) 2-57 Isomer 2 6.0 (40) <4.7(11) <4.7 (23)  7.4 (107) 2-58 Isomer 2 5.9 (63) NT NT 6.6 (74) 2-59Isomer 2 <4.7 (9)    <4.7 (9)  <4.7 (14) 7.1 (52) 2-60 Isomer 2 <4.7(62)   <4.7 (20) <4.7 (56)  7.2 (111) 2-61 Isomer 2 <4.7 (21)   <4.7(52) <4.7 (65)  7.4 (101) 2-62 Isomer 2 6.0 (85) <4.7 (12) <4.7 (11) 7.7 (106) 2-63 Isomer 2b <4.7 (7)    NT NT 6.2 (86) 2-64 Isomer 2 5.6(46) <4.7 (2)  <4.7 (4)   7.5 (128) 2-65 Isomer 1a 5.1 (45) NT NT <4.7(14)   2-65 Isomer 2a 6.0 (43) <4.7 (15) <4.7 (15) 6.7 (45) 2-66 Isomer2 <4.7 (59)   <4.7 (17) <4.7 (8)  7.4 (76) 2-67 Isomer 2 <4.7 (19)  <4.7 (2)  <4.7 (2)  7.5 (55) 3-1 Racemic <4.7 (19)   <4.7 (2)  <4.7 (5) 8.0 (46) 3-2 Isomer 2 6.5 (42) <4.7 (29) <4.7 (37) 7.5 (94) 3-3 Isomer 2<4.7 (18)   <4.7 (3)  <4.7 (21) 8.4 (93)

Example B

Effect of a Novel Test Compound and Xanomeline on d-Amphetamine-InducedHyperactivity in Rats

The aim of the study is to examine the effect of a novel test compoundon d-amphetamine induced hyperactivity in rats. Schizophrenia is acomplex multifactorial disease that cannot be fully represented by asingle experimental procedure. Antipsychotic-like behaviour was assessedin rats by the inhibition of hyperactivity (or hyperlocomotion) elicitedby d-amphetamine. This procedure is sensitive to clinically relevantdopamine receptor antagonists and is therefore considered suitable forcomparing muscarinic agonists that influence dopaminergic signalling. Adose of xanomeline previously observed to significantly reduced-amphetamine induced hyperactivity was employed as a positive control.Statistical analysis typically involved three-way analysis of covarianceor robust regression with treatment, day and rack as factors andactivity during the 30 minutes prior to treatment as a covariate,followed by appropriate multiple comparison tests. A P value of <0.05was considered statistically significant and is marked accordingly inall subsequent figures.

Data for Example 2-1 Isomer 2 is shown in FIG. 1.

Example C

Pharmaceutical Formulations

(i) Tablet Formulation

A tablet composition containing a compound of the formula (1) or formula(1a) is prepared by mixing 50 mg of the compound with 197 mg of lactose(BP) as diluent, and 3 mg magnesium stearate as a lubricant andcompressing to form a tablet in known manner.

(ii) Capsule Formulation

A capsule formulation is prepared by mixing 100 mg of a compound of theformula (1) or formula (1a) with 100 mg lactose and optionally 1% byweight of magnesium stearate and filling the resulting mixture intostandard opaque hard gelatin capsules.

EQUIVALENTS

The foregoing examples are presented for the purpose of illustrating theinvention and should not be construed as imposing any limitation on thescope of the invention. It will readily be apparent that numerousmodifications and alterations may be made to the specific embodiments ofthe invention described above and illustrated in the examples withoutdeparting from the principles underlying the invention. All suchmodifications and alterations are intended to be embraced by thisapplication.

The invention claimed is:
 1. A compound of the formula (2):

or a salt thereof, wherein q is 1; r is 1 or 2; s is 0 or 1, where thetotal of r and s is 1 or 2; R⁴ is a hydrogen or a C₁₋₆ non-aromatichydrocarbon group which is optionally substituted with one to sixfluorine atoms; R⁵ is selected from a C₁₋₆ non-aromatic hydrocarbongroup which is optionally substituted with one to six fluorine atoms andwherein one or two, but not all, carbon atoms of the hydrocarbon groupmay optionally be replaced by a heteroatom selected from O, N and S andoxidized forms thereof; and a group W or CH₂W where W is an optionallysubstituted 5- or 6-membered aromatic ring containing 0, 1, 2 or 3heteroatoms selected from O, N and S; R⁶ is selected from; COR⁷; COOR⁷;CH₂COR⁷; CH₂COOR⁷ and a C₁₋₆ non-aromatic hydrocarbon group which isoptionally substituted with one to six fluorine atoms and wherein one ortwo, but not all, carbon atoms of the hydrocarbon group may optionallybe replaced by a heteroatom selected from O, N and S and oxidized formsthereof; wherein R⁷ is selected from H and a C₁₋₆ non-aromatichydrocarbon group which is optionally substituted with one to sixfluorine atoms.
 2. The compound according to claim 1 wherein r is 1 ands is
 1. 3. The compound according to claim 1 wherein r is 1 or 2 and sis
 0. 4. The compound according to claim 1 wherein R⁴ is selected fromhydrogen and methyl.
 5. The compound according to claim 1 wherein R⁵ isselected from methyl, ethyl, propyl, isopropyl, cyclopropyl,fluoroethyl, difluoroethyl, butyl and cyclobutyl.
 6. The compoundaccording to claim 1 wherein R⁵ is a group W or CH₂W where W is anoptionally substituted phenyl, pyridyl or isoxazole ring.
 7. Thecompound according to claim 1 wherein R⁶ is selected from methyl, ethyl,trifluoroethyl, hydroxyethyl and methoxyethyl.
 8. The compound accordingclaim 1 wherein R⁶ is selected from; COR⁷; COOR⁷; CH₂COR⁷; and CH₂COOR⁷,wherein R⁷ is selected from H, methyl, fluoromethyl, difluoromethyl,trifluoromethyl, ethyl, fluoroethyl, difluoroethyl and trifluoroethyl.9. The compound according to claim 1 which is Ethyl6-{4-[acetyl(cyclopropyl)amino]piperidin-1-yl}-2-azaspiro[3.3]heptane-2-carboxylate;Ethyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.3]heptane-2-carboxylate;Ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(propan-2-yl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(cyclopropyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(trifluoroacetyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(propanoyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(cyclobutyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(2-methoxyethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(2-methoxy-2-oxoethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(1,2-oxazol-3-ylmethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Methyl2-{4-[acetyl(cyclobutyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(formyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Methyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(propyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(2-methylpropyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(cyclopropylmethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-(4-{acetyl[(1-methylcyclobutyl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(1-methylcyclobutyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-(4-{acetyl[(3-methyloxetan-3-yl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(2,2-difluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(2-fluoro-2-methylpropyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(2,2-difluoropropyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(3,3-difluorocyclobutyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(2-methoxy-2-methylpropyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-(4-{acetyl[(1-methoxycyclobutyl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-(4-{acetyl[(1-hydroxycyclobutyl)methyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(methoxy)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(phenyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(pyridin-2-yl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(benzyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(1,3-oxazol-5-ylmethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[acetyl(1,3-oxazol-2-ylmethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;(1,1-²H₂)-Ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(methoxycarbonyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(ethoxycarbonyl)(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(methoxycarbonyl)(propan-2-yl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(methoxycarbonyl)(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(2-fluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(2,2-difluoroethyl)(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(methoxycarbonyl)(oxetan-3-yl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclopropyl(2-hydroxyethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[cyclobutyl(2-hydroxyethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(2-hydroxyethyl)(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(2-methoxyethyl)(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(oxetan-3-yl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[oxetan-3-yl(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(2-cyanoethyl)(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-(4-{ethyl[2-(methylsulfonyl)ethyl]amino}piperidin-1-yl)-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(methoxy)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(2-hydroxyethyl)(phenyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[benzyl(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[ethyl(1,3-oxazol-5-ylmethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(2-fluoroethyl)(1,3-oxazol-5-ylmethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl2-{4-[(1,3-oxazol-5-ylmethyl)(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylate;Ethyl6-{4-[acetyl(cyclopropyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylate;Methyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylateor Ethyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylate,or a salt of any of the foregoing.
 10. The compound according to claim 1which is ethyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.3]heptane-2-carboxylateor a pharmaceutically acceptable salt thereof.
 11. The compoundaccording to claim 1 which is ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 12. The compoundaccording to claim 1 which is ethyl2-{4-[ethyl(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 13. The compoundaccording to claim 1 which is methyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 14. The compoundaccording to claim 1 which is ethyl2-{4-[acetyl(2,2-difluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 15. The compoundaccording to claim 1 which is (1,1-²H₂)-ethyl2-{4-[acetyl(ethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 16. The compoundaccording to claim 1 which is ethyl2-{4-[(2-methoxyethyl)(2,2,2-trifluoroethyl)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 17. The compoundaccording to claim 1 which is ethyl2-{4-[ethyl(methoxy)amino]piperidin-1-yl}-6-azaspiro[3.4]octane-6-carboxylateor a pharmaceutically acceptable salt thereof.
 18. The compoundaccording to claim 1 which is methyl6-{4-[acetyl(ethyl)amino]piperidin-1-yl}-2-azaspiro[3.4]octane-2-carboxylateor a pharmaceutically acceptable salt thereof.
 19. A pharmaceuticalcomposition comprising a compound as defined in claim 1 and apharmaceutically acceptable excipient.
 20. The compound according toclaim 1 wherein the compound has a muscarinic M₁ receptor and/or M₄receptor agonist activity.